FROM ALGAE IN EDIBLE PRODUCTS

CROSS-REFERENCE TO RELATED APPLICATION(S)

[0001] This application claims the benefit of priority under 35 U.S.C. § 119(e) of US Provisional Application No. 62/865,800, filed June 24, 2019, of US Provisional Application No. 62/850,227, filed May 20, 2019, and of US Provisional Application No. 62/757,534, filed November 8, 2018, the entire content of each of which is hereby incorporated by reference.

SEQUENCE LISTING

[0002] The instant application contains a Sequence Listing which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on November 6, 2019, is named 20498-202379_SL.txt and is 208 kilobytes in size.

BACKGROUND

[0003] With the advent of industrialized animal agriculture, the consumption of animal meat has continued to rise. Animal agriculture requires a significant amount of land use and fresh water, finite resources that are becoming increasingly difficult to access.

[0004] To address the sustainability and ethical concerns over animal meat consumption, the food industry has been aggressively trying to develop plant-based alternatives that taste, touch and smell like meat products. However, many of the current plant-based alternatives have not been able to penetrate the larger food and consumer markets. To improve the sustainability of the food ecosystem it is imperative that products are developed that appeal to consumers who currently prefer meat.

[0005] Recent advances made have demonstrated the potential of using heme-containing proteins, purified from a host organism, to make the flavor and aroma profile of a product closer to that of meat. It is thought that the heme from heme-containing proteins are responsible for imparting a“meaty” flavor and aroma to meat products. However, the available sources of heme-containing proteins are expensive and technically intensive limiting their utility. In addition to poor economics, the product is genetically modified making it less appealing to many consumers who have chosen to consume foods that are not a result of genetic engineering. Additionally, there is a trend to products with increased nutrition benefit and a balance of caloric intake. A number of the current meat alternatives cannot fully satisfy these demands while maintain the taste, texture and visual appeal desired by consumers. Thus, a need exists for edible products incorporating heme-containing proteins as set forth herein.

SUMMARY OF THE INVENTION

[0006] To address both the economic and consumer concerns associated with the current approaches of incorporating heme into a product, provided herein are compositions and processes for producing such compositions that provide flavorful and nutritious alternatives to meat. In particular, provided herein are compositions and methods of producing such compositions that incorporate heme from algae, along with other nutrition components. Algae can be incorporated into finished products without the costly process of purification.

[0007] The present disclosure includes compositions of engineered algae overexpressing or accumulating heme and methods of using such engineered algae for food products. Thus, one aspect of the disclosure includes an engineered algae having a genetic modifications, where the genetic modification results in an accumulation of heme in the algae as compared to an algae lacking the genetic modification. In some embodiments, the engineered algae has reduced or absence of chlorophyll production. In some embodiments, the algae has red or red-like color. In some embodiments, the algae is capable of growth on glucose as the sole carbon source.

[0008] Preferably, the genetic modification comprises a genetic alteration to chlorophyll synthesis pathway, protoporphyrinogen IX synthesis pathway or heme synthesis pathway. In some embodiments, the genetic modification is associated with a deficiency in the expression of magnesium chelatase. Alternatively and/or additionally, the genetic modification comprises an alteration in one or more of CHLD, CHLI1, CHLI2 or CHLH1. Alternatively and/or additionally, the genetic modification comprises an alteration in an upstream regulatory region, a downstream regulatory region, an exon, an intron or any combination thereof. In some embodiments, the genetic modification comprises an insertion, a deletion, a point mutation, an inversion, a duplication, a frameshift or any combination thereof.

[0009] In some embodiments, the engineered algae has a heme content greater than the chlorophyll content. Alternatively and/or additionally, the engineered algae has a

protoporphyrin IX content greater than the chlorophyll content. Alternatively and/or additionally, the engineered algae has reduced production of one or more fatty acids. [0010] In some embodiments, the engineered algae further comprises a genetic modification that reduces or eliminates the expression of light independent

protochlorophyllide oxidoreductase. In such embodiments, it is contemplated that the genetic modification comprises a mutation or deletion in one or more of ChlB, ChlL or ChlN. In some embodiments, the engineered algae has upregulated expression of ferrocheletase and/or upregulated expression of protoporphyrinogen IX oxidase. Optionally, the algae contain a recombinant or heterologous nucleic acid. In some embodiments, the engineered algae comprises a Chlamydomonas sp. Alternatively and/or additionally, the Chlamydomonas sp. is Chlamydomonas reinhardtii.

[0011] Another aspect of the disclosure includes an edible composition comprising an algae preparation, wherein the algae preparation comprises an engineered algae as described above or a portion thereof. In some embodiments, the edible composition comprises heme derived from the engineered algae. In some embodiments, the algae preparation comprises algae cells. In some embodiments, the algae preparation is a fractionated algae preparation. In some embodiments, the algae preparation is red or red-like in color.

[0012] In some embodiments, the edible composition has a red or red-like color derived from the algae preparation. Alternatively and/or additionally, the algae preparation confers a meat or meat-like flavor to the edible composition. Alternatively and/or additionally, the edible composition has a meat or meat-like texture derived from the algae preparation. In such embodiment, it is contemplated that the meat or meat-like texture comprises a beef or beef-like texture, a fish or fish-like texture, a chicken or chicken-like texture, a pork or pork- like texture or a texture of a meat replica.

[0013] In some embodiments, the edible composition is a finished product selected from the group consisting of a beef-like food product, a fish-like product, a chicken-like product, a pork-like product and a meat replica. Alternatively and/or additionally, the edible composition is vegan, vegetarian or gluten-free. Alternatively and/or additionally, the edible composition has an appearance of blood derived from the algae preparation.

[0014] Alternatively and/or additionally, the algae preparation has a heme content greater than the chlorophyll content. Alternatively and/or additionally, the algae preparation has a protoporphyrin IX content greater than the chlorophyll content. In some embodiments, the algae preparation provides at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or 100% of the total protein content to the edible composition. Alternatively and/or additionally, the algae preparation provides vitamin A, beta carotene or a combination thereof to the composition. Optionally, the vitamin A, the beta carotene or the combination thereof is at least about 5%, 10%, 15%,

20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%,

99% or 100% of the daily recommended requirement. Alternatively and/or additionally, the algae preparation provides less than about 0.01%, 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.2%, 1.5%, 2%, 5% or 10% of total saturated fat present in the edible composition. Alternatively and/or additionally, the algae preparation provides less than about 0.01%, 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%,

1.2%, 1.5%, 2%, 5% or 10% of total saturated fat present in a finished product comprising the edible composition. Alternatively and/or additionally, the algae preparation provides at least about 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg, 95 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg or 500 mg of omega-3 fatty acids to the edible composition. Alternatively and/or additionally, the algae preparation has reduced fatty acid content.

[0015] In some embodiments, the edible product is combined with a protein source, a fat source, a carbohydrate, a starch, a thickener, a vitamin, a mineral, or any combination thereof. In such embodiments, it is preferred that the protein source is selected from the group consisting of textured wheat protein, textured soy protein and textured pea protein, fungal protein or algal protein. Alternatively and/or additionally, the fat source comprises at least one of refined coconut oil or sunflower oil. In some embodiments, the edible component further comprises at least one of potato starch, methylcellulose, water, and a flavor, wherein the flavor is selected at least one of yeast extract, garlic powder, onion powder, and salt.

[0016] In some embodiments, the edible product is an ingredient for a burger, a sausage, a kebab, a filet, a fish-altemative, a ground meat-like product or a meatball. In some embodiments, the burger comprises about 5% of the algae preparation, about 20% textured soy protein and about 20% refined coconut oil. Optionally, the burger further comprises about 3% sunflower oil, about 2% potato starch, about 1% methylcellulose, about 45% water and about 4-9% flavors. Alternatively and/or additionally, the burger further comprises about 0.5% Kojac gum, about 0.5% Xanthan gum, about 45% water and about 4-9% flavors. In some embodiments, fish-altemative comprises 20% textured soy protein, about 5 % of algae preparation, about 65% water and about 10% flavors. In some embodiments, the edible composition is free of animal proteins.

[0017] In some embodiments, the algae preparation comprises an algae having an increase in protoporphyrinogen IX synthesis or accumulation. Alternatively and/or additionally, the algae preparation comprises an algae that exhibits a red or red-like color when grown in the dark conditions. In some embodiments, the algae comprised in the algae preparation are recombinant or genetically modified algae. In some embodiments, the algae preparation comprises a Chlamydomonas sp. Optionally, the Chlamydomonas sp. is

Chlamydomonas reinhardtii.

[0018] Another aspect of the disclosure includes a method for the production of an edible composition. The method includes steps of (a) culturing an engineered algae as described above in a condition where the engineered algae exhibits a red or red-like color and wherein the engineered algae produces heme, (b) collecting the cultured engineered algae to produce an algae preparation, and (c) combining the algae preparation with at least one edible ingredient to produce an edible composition. In some embodiments, the condition comprises a fermentation condition. Alternatively and/or additionally, the condition comprises acetate as a reduced carbon source for growth of the engineered algae. Alternatively and/or additionally, the condition comprises sugar as a reduced carbon source for growth of the engineered algae. Alternatively and/or additionally, the condition comprises dark or limited light condition. Alternatively and/or additionally, the condition further comprises iron supplements.

[0019] In some embodiments, the method further comprises fractionating the cultured algae to produce the algae preparation. In some embodiments, the algae preparation has a heme content that is greater than the chlorophyll content. Alternatively and/or additionally, algae preparation has a protoporphyrin IX content that is greater than the chlorophyll content. In some embodiments, the engineered algae is a Chlamydomonas sp. Optionally, the engineered algae is a Chlamydomonas reinhardtii.

[0020] In some embodiments, the edible composition has at least one of the features selected from the group consisting of a meat or meat-like flavor, a meat or meat-like texture, a blood-like appearance and a meat or meat-like color, where the at least one of the features is derived from the algae preparation. In some embodiments, the method further comprises producing a finished product comprising the edible composition and wherein the finished product is a beef-like food product, a fish-like product, a chicken-like product, a pork-like product or a meat replica. In some embodiments, the edible composition is free of animal proteins. In some embodiments, the algae preparation is fractionated to remove one or more of starch, protein, PPIX, fatty acids and chlorophyll.

[0021] Another aspect of the disclosure includes a method of making an engineered algae enriched in heme content. The method includes steps of (a) subjecting an algae strain to a process that produces genetic modification to create a first algae population, and (b) from the first algae population, selecting a second algae population that is enriched in heme content, and optionally, PPIX content. In some embodiments, the process comprises at least one of a random UV mutagenesis, a random chemical mutagenesis, a recombinant genetic engineering, a gene editing, or a gene silencing. In some embodiments, the method further comprises a step of culturing the first algae population in a fermentation condition. In some embodiments, the fermentation condition comprises a media having sugar as a sole carbon source. In such embodiments, it is preferred that the sugar is selected from glucose, dextrose, fructose, maltose, galactose, sucrose, and ribose. Alternatively and/or additionally, the fermentation condition comprises a brightness of less than 500 lux.

[0022] In some embodiments, the selecting the second algae population step comprises sorting or identifying algae cells having a red or red-like color. Alternatively and/or additionally, the second algae population step is performed by FACS. In some embodiments, the second algae population is selected with its capability to grow in the fermentation condition.

BRIEF DESCRIPTION OF THE DRAWINGS

[0023] FIG. 1 is a pictorial diagram showing an exemplary pathway for the production of heme in algae. This exemplary pathway can be used by wildtype algae to produce chlorophyll, but it can also be used to generate heme.

[0024] FIGs. 2A and 2B show the composition of an exemplary algae growth media (Fig. 2A) and selection process (Fig. 2B).

[0025] FIG. 3 is a pictorial diagram showing algae growth in complete dark condition with dextrose as the only carbon source. [0026] FIG. 4 is a pictorial diagram showing an exemplary fractionation of algae overexpressing heme, showing the separation into a protein and heme-enriched biomass, which is separated from the starch and carotenoid fractions.

[0027] FIG. 5 is a pictorial diagram showing extraction process of PPIX and/or heme from the red algae.

[0028] FIG. 6 is a graphical diagram showing an exemplary growth curve (dry cell weight) of a heme-overproducing strain when grown in aerobic fermentation conditions.

[0029] FIG. 7 is a graphical diagram showing increased dry cell weight of

Chlamydomonas sp. in a glucose-containing media.

[0030] FIG. 8 is a graphical diagram showing the fractionated components of the red algae preparation before and after hexane extraction.

[0031] FIG. 9 shows a portion of sequence alignments of a wild type green algae and a red-algae with a mutation in CHLH gene (upper sequence (Seq l) is a partial nucleic acid sequence (residues 1621-1679 of SEQ ID NO: 27) and a partial amino acid sequence (residues 451-460 of SEQ ID NO: 28) of CHLH gene of green algae, and lower sequence (Seq_2) is a partial nucleic acid sequence (residues 1621-1680 of SEQ ID NO: 129) and partial amino acid sequence (residues 451-460 of SEQ ID NO: 152) of CHLH gene of red algae has a mutation (asterisk)). As shown, the wild-type CHLH nucleic acid sequence (SEQ ID NO: 27) has an insertion of a thiamine at position 1678 resulting in a change of the wild- type CHLH amino acid sequence of SEQ ID NO: 28 of a proline to a serine at amino acid position 560.

[0032] FIG. 10 is a pictorial diagram showing burgers created with O.Olg, 0.1 g, l.Og, and 5.0g of the heme enriched algae.

[0033] FIG. 11 is a pictorial diagram showing ingredient mixes of the plant-based burger ingredients with no heme-enriched algae, with the addition of heme-enriched algae, or the ingredients with the addition of heme-enriched algae shaped into a burger before and after cooking.

[0034] FIG. 12 is a pictorial diagram showing an example of heme-enriched meatless “tuna”. DETAILED DESCRIPTION

[0035] Before the present compositions and methods are described, it is to be understood that this invention is not limited to particular compositions, methods, and experimental conditions described, as such compositions, methods, and conditions may vary. It is also to be understood that the terminology used herein is for purposes of describing particular embodiments only, and is not intended to be limiting, since the scope of the present invention will be limited only in the appended claims.

[0036] As used in this specification and the appended claims, the singular forms“a”,“an”, and“the” include plural references unless the context clearly dictates otherwise. Thus, for example, references to“the method” includes one or more methods, and/or steps of the type described herein which will become apparent to those persons skilled in the art upon reading this disclosure and so forth. Furthermore, to the extent that the terms“including”,“includes”, “having”,“has”,“with”, or variants thereof are used in either the detailed description and/or the claims, such terms are intended to be inclusive in a manner similar to the term

“comprising.”

[0037] The term“about” or“approximately” means within an acceptable error range for the particular value as determined by one of ordinary skill in the art, which will depend in part on how the value is measured or determined, e.g., the limitations of the measurement system. For example,“about” can mean within 1 or more than 1 standard deviation, per the practice in the given value. Where particular values are described in the application and claims, unless otherwise stated the term“about” should be assumed to mean an acceptable error range for the particular value.

[0038] As used herein,“a deficiency in” or the“lack of’, or“reduction of’, one or more genes and/or enzymes include, for example, mutation or deletion of the gene sequence, a reduction in or lack in the expression from a gene (RNA and/or protein) and/or a lack of accumulation or stability of a gene product (RNA and/or protein).

[0039] As used herein,“overexpresses” and“overexpression” of an enzyme or gene include, for example, an increase in expression from a gene (RNA and/or protein) and/or an increase in accumulation or stability of a gene product (RNA and/or protein). Such overexpression can include alterations to the regulatory region(s) and/or to the gene sequence, as well as copy number, genomic position and post-translational modifications. [0040] As used herein, the term“engineered algae” is used to refer to an algae that contains one or more genetic modifications. In some cases, an engineered algae is also a recombinantly modified organism when it incorporates heterologous nucleic acid into its genome through recombinant technology. In other cases, an engineered algae is not a recombinantly modified organism (for example when it is modified through UV, chemical or radiation mutagenesis). In some cases an algae that is not a recombinantly modified organism is referred to as non-GMO, and components from such algae can be referred to as non-GMO components.

[0041] As used herein, the term“genetic modification” is used to refer to any

manipulation of an organism’s genetic material in a way that does not occur under natural conditions. A genetic modification can include modifications that are made through mutagenesis (such as with UV light, X-rays, gamma irradiation and chemical exposure). A genetic modification can include gene editing. In some cases, genetic modifications can be made through recombinant technology. As used herein,“recombinantly modified organism” is used to refer to an organism that incorporates heterologous nucleic acid (e.g., recombinant nucleic acid) into its genome through recombinant technology. Methods of performing such manipulations are known to those of ordinary skill in the art and include, but are not limited to, techniques that make use of vectors for transforming cells with a nucleic acid sequence of interest. Included in the definition are various forms of gene editing in which DNA is inserted, deleted or replaced in the genome of a living organism using engineered nucleases, or“molecular scissors.” These nucleases create site-specific double-strand breaks (DSBs) at desired locations in the genome. The induced double-strand breaks are repaired through nonhomologous end-joining (NHEJ) or homologous recombination (HR), resulting in targeted mutations (i.e.. edits).

[0042] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the invention, the preferred methods and materials are now described.

[0043] Provided herein are compositions and methods to provide heme and other nutrition components from algae. Algae are known for producing many compounds that result in these aquatic organisms being various colors. These compounds include, but are not limited to, chlorophyll which makes algae green, beta-carotene which makes algae appear yellow or orange, astaxanthin which makes algae appear red or other various pigments such as phycocyanin which make algae blue. While each of these previously mentioned compounds has been added to food products, there are to date no products that incorporate an algae over producing heme to impart a red color and/or a meaty taste and smell.

[0044] Provided herein are strains, methods and compositions that employ algae overproducing heme. In some embodiments, the algae strain when grown is red or red-like in color. As used herein, in some embodiments, red-like color can be any color with a wavelength between 590 nm to 750 nm or any mixture of the color. Alternatively and/or additionally, in some embodiments, red-like color can be defined as any color in RGB (r.g.b) having r value between 255 and 80 with g or b values between 0 and 80. In some

embodiments, a preparation made from the algae culture overproducing heme, imparts a pink or red color when incorporated into food and other edible products. In some embodiments, a preparation made from the algae culture overproducing heme, imparts a“meaty” flavor, smell and/or texture when incorporated into food and other edible products. In some embodiments, a preparation made from the algae culture overproducing heme, imparts a desired color, taste and/or smell, as well as one or more additional nutrition components such as omega-3 fatty acids, saturated fats, protein, vitamin A, beta-carotene or any combination thereof.

Algae Producing and Over-Producing Heme

[0045] Provided herein are algae strains that over-produce heme and strains that produce or accumulate heme and/or protoporphyrin IX (PPIX) content greater than chlorophyll content and that can be used to produce edible compositions and ingredients. Also provided herein are methods of making such strains and ingredients and compositions therefrom and use with the methods herein to make such compositions. Such strains are created by modifying one or more steps in the biochemical pathways that produce heme, PPIX and chlorophyll.

[0046] Without being bound by theory, the heme pathway is a biochemical pathway that branches from the chlorophyll biochemical pathway, as shown in FIG. 1. In short, this pathway starts with a glutamate tRNA which is converted to 5-aminolaevulinic acid (ALA) by a GlutRNA reductase and a GSA amino transferase. Next, ALA is converted to porphobilinogen by ALA dehydrase. Next, porophobilinogen is converted to hydroxymethylbilane by pophobibnogen deaminase. Next, hydroxymethylbilane is converted to uroporphyrinogen III by UPG III synthase. Next, uroporphyrinogen III is converted to coprophyrinogen by UPG III decarboxylase. Next, coprophyrinogen is converted to protoporphyrinogen IX by CPG oxidase. Next, protoporphyrinogen IX is converted to protoporphyrin IX by PPG oxidase. Protoporphyrin IX can be shutled to the chlorophyll production pathway or towards heme B. Finally, protoporphyrin IX is converted to heme B by the enzyme ferrochelatase which ataches iron to protoporphyrin IX.

[0047] By reducing metabolic flux towards chlorophyll, it is possible to increase metabolic flux towards heme B. In some embodiments herein, the algae strains used in the methods and compositions produced therewith are reduced in metabolic flux towards chlorophyll and increased metabolic flux towards heme B (also referred to herein as“heme”). In some embodiments, the algae strain is one where chlorophyll and carotenoid synthesis is decreased and heme synthesis or accumulation is increased. In some embodiments, the algae strain is deficient or reduced in the amount of chlorophyll. In some embodiments, the algae strain is red or red-like in color.

[0048] In some embodiments, the algae strain is deficient for one or more enzymes in the chlorophyll biosynthesis pathway. Such deficiencies include, but are not limited to, gene deletions, mutations and other alterations that result in a lack expression of the enzyme or a deficiency in the functionality of the enzyme. In some embodiments, the algae strain is deficient in magnesium chelatase which is the first step in converting protoporphyrin IX to chlorophyll. In some embodiments, the algae strain is deficient for light dependent protochlorophyllide which converts protochlorophyllide to chlorophyllide. In some embodiments, the algae strain is deficient for a light independent protochlorophyllide which converts protochlorophyllide to chlorophyllide in the dark. In some embodiments, the algae strain is deficient for one or more of ChlB, ChlL, or ChlN gene products which are encoded in the chloroplast genome and are subunits of light independent protochlorophyllide oxidoreductase (LIPOR) that coverts protochlorophyllide to chlorophyllide. This enzyme, when expressed, can allow algae such as Chlamydomonas to produce chlorophyll and remain green even when the algae is not provided with illumination. When one or more of these genes are knocked out, the algae strain has a yellow color under dark growing conditions. [0049] In some embodiments, the algae strain is lacking or reduced in one or more of magnesium chelatase, magnesium protoporphyrinogen IX, protochlorophyllide,

chlorophyllide, and chlorophyll.

[0050] In some embodiments, the algae strain is deficient for one or more of the magnesium chelatase subunits CHLD, CHLH and CHLI. These subunits are also referred to by the gene names, CHLD 1 (alternatively written as CH1D1), corresponding to the CHLD subunit, CHLH1 (alternatively written as CH1H1), corresponding to the CHLH subunit, and CHLI1 and CHLI2, corresponding to the CHLI subunit, encoded by two genes, CHLH and CHLI2 (alternatively written as CH1I1 and CH1I2).

[0051] In some embodiments, a heme-enriched algae strain is deficient in one or more of a nuclearly encoded subunit of magnesium chelatase, for example in one or more of the subunits encoded by the genes for the subunits CHLD, CHLH and CHLI. A deficiency in one or more of these subunits reduces or eliminates chlorophyll expression. In some embodiments, the gene encoding a subunit can be modified, such as by one or more point mutations that change a codon to a stop codon, resulting in a truncated coding region. In some embodiments, the gene encoding a subunit can be modified by a deletion that removed some of or all of the gene encoding the subunit. In some embodiments, the gene encoding a subunit can be modified by a frameshift mutation, such as caused by a deletion or insertion of one or more bases into the coding region, resulting in a non-functional and/or truncated protein. In some embodiments, the gene encoding a subunit can be modified by an insertion into the coding region that creates a non-functional protein, such as by adding one or more amino acids internally or at the N or C terminus of the protein that creates a non-functional subunit or reduces the activity or stability of the subunit or enzyme.

[0052] In some embodiments, a heme-enriched algae has at least one modification in the nucleotide sequence encoding CHLD, CHLH, CHLI2 or CHLH1 (e.g., a modification in SEQ ID NOs: 23, 25, 27, 153) including the intron, exon, regulatory regions, or full gene sequences. In some embodiments, a heme-enriched algae has at least one modification in the amino acid sequence of CHLD, CHLH, CHLI2 or CHLH1 (e.g., a modification in SEQ ID NOs: 24, 26, 28, 151). In some embodiments, a heme-enriched algae strain contains at least one modification (point mutation, deletion, or insertion) in an exon encoding a portion of CHLD, CHLH, CHLI2 or CHLH1. In some embodiments, a heme-enriched algae strain contains at least one modification to a wildtype sequence of such exons, such as a modification in any of SEQ ID NOs: 47-58, 72-80, 91-102, and 132-141.

[0053] In some embodiments, a heme-enriched algae strain contains at least one modification (point mutation, deletion, or insertion) in an untranslated region of CHLD, CHLI1, CHLI2 or CHLH1, such as in the 5’ untranslated region or the 3’ untranslated region. In some embodiments, a heme-enriched algae strain contains at least one modification to a wildtype sequence of such untranslated regions, such as a modification in any of SEQ ID NOs: 45, 46, 70, 71, 89, 90, 130 or 131.

[0054] In some embodiments, the regulation of expression of one or more subunit of Mg- chelatase is altered to create a strain that has reduced amounts of chlorophyll. The regulatory regions of one or more of CHLD, CHLI1, CHLI2 and CHLH1 can be modified to reduce expression, such as by an insertion, deletion or one or more point mutations. Such alterations may modify, for example, transcription factor binding sites, enhancer sites, RNA polymerase interactions and transcriptional start sites in a manner the reduces or eliminates the transcription of a subunit gene.

[0055] In some embodiments, the expression of one or more subunits is altered by modifying the splicing of an intron with the gene of a subunit, such as a mutation, insertion or deletion that eliminates or alters a splicing donor or acceptor site or that otherwise alters the efficiency or accuracy of the gene splicing. In some embodiments, a heme-enriched algae strain contains at least one modification (point mutation, deletion, or insertion) in an intron of CHLD, CHLI1, CHLI2 or CHLH1. In some embodiments, a heme-enriched algae strain contains at least one modification to a wildtype sequence of such introns, such as a modification in any of SEQ ID NOs: 59-69, 81-88, 103-113, 142-150.

[0056] In some embodiments, the algae strain overexpresses one or more enzymes such that the balance of pathways favors heme production. In some embodiments, the algae strain overexpresses one or more of glutamyl-tRNA reductase, glutamyl- 1 -semialdehyde aminotransferase, ALA dehydrongenase, porphobilinogen deaminase, UPG III synthase,

UPG III decarboxylase, CPG oxidase, PPG oxidase, and ferrochelatase. In some

embodiments, the algae strain is improved for its ability to produce ALA, a rate limiting precursor of heme B synthesis. In some embodiments, the algae strain is improved for its ability to produce a functional ferrochelatase gene, the enzyme responsible for the conversion of protoporphyrin IX to heme B. In some embodiments, the algae strain is improved for its ability to produce UPG III synthase, UPG III decarboxylase, CPG oxidase, or PPG oxidase. In some embodiments, the algae strain has an increased amount of one or more of heme, a heme-containing protein, protoporphyrinogen IX, biliverdin IX, photochromobilin, and ferrocheletase, as compared to a wildtype strain.

[0057] In some embodiments, the algae strain produces carotenoids or precursors of carotenoids. Without being bound by theory, carotenoids confer color and can have an impact on the visual appearance of a plant-based alternative. Exemplary carotenoids include, but are not limited to, gamma-carotene, beta-carotene, beta cryptoxanthin, zeaxanthin, autheraxanthin, lutein, prolycopene and lycopene.

[0058] In some embodiments, the algae strain is deficient for carotenoids or precursors of carotenoids. Deficiencies in carotenoid biosynthesis can occur due to mutations, such as mutations that impact carotenoid biosynthesis, for example, mutations in the phytoene synthase gene.

[0059] In some embodiments herein, algae used in the compositions and methods herein is non-GMO, does not contain heterologous nucleic acid and/or is not created using recombinant technology. In some embodiments, algae used in the compositions and methods herein is selected based on its color, heme content, rate of heme synthesis, accumulation of heme, or protoporphyrin IX content, rate of synthesis or accumulation. In some

embodiments, the algae have reduced levels of chlorophyll and/or levels of chlorophyll that are less than the levels of heme and/or protoporphyrin IX. In some embodiments, algae used in the compositions and methods herein does not contain a heterologous gene for one more genes involved in heme biosynthesis or accumulation, e.g., the algae does not contain a bacterial, fungal, plant or animal-derived gene or nucleic acid that is involved in heme biosynthesis, heme accumulation, protoporphyrin IX biosynthesis, or protoporphyrin IX accumulation.

[0060] In some embodiments, algae are modified in expression of one or more genes contributing to an increase in heme synthesis or accumulation, a decrease in chlorophyll synthesis or accumulation or a combination thereof. Such modifications can be created through mutagenesis such as by exposure to UV light, radiation or chemicals. [0061] In some embodiments, modifications can be created through gene editing such as precisely engineered nuclease targeting to alter the expression of one or more components, such as by CRISPR-CAS nucleases. Such nucleases can be used to create insertions, deletions, mutations and replacements of one or more nucleotides or regions of nucleotides to modify the expression of one or more pathway enzymes in the pathway to reduce chlorophyll and/or to increase the production of heme. Subsequent to the creation of the modification, the algae strain can be grown and/or mated such that the nuclease and associated guide nucleic acids are removed, and the algae strain that remains does not retain the nuclease and associated editing system. In some embodiments, a nuclease such as the CRISPR-CAS nuclease can be used to make a modification to a component of the chlorophyll pathway such that chlorophyll expression and/or accumulation is reduced or abrogated. In some embodiments, a nuclease such as the CRISPR-CAS nuclease can be used to make a modification to a component of the chlorophyll pathway such that heme expression and/or accumulation is increased. In some embodiments, a nuclease such as the CRISPR-CAS nuclease is used to make a modification in one or more of CHLD, CHLI1, CHLI2 or CHLH1 resulting in a heme-enriched algae strain. Such modifications can be made by designing guide RNAs with modifications to one or more of SEQ ID NOs:45-l l3, 130-150 and/or 153 to include one or more point mutations, insertions, deletions or combinations thereof.

[0062] There are several families of engineered nucleases that can be used for gene editing described herein, for example, but not limited to, meganucleases, zinc finger nucleases (ZFNs), transcription activator-like effector-based nucleases (TALEN), the CRISPR-Cas system, and ARCUS. However, it should be understood that any known gene editing system utilizing engineered nucleases may be used in the methods described herein. Thus, in some embodiments, the algae strain overproducing heme can be created by using techniques such as a CRISPR-Cas system (e.g., CRISPR-CAS9) or by the use of zinc-finger nucleases.

[0063] CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) is an acronym for DNA loci that contain multiple, short, direct repetitions of base sequences. The prokaryotic CRISPR/Cas system has been adapted for use as gene editing (silencing, enhancing or changing specific genes) for use in eukaryotes (see, for example, Cong,

Science, 15:339(6121):819-823 (2013) and Jinek, et al, Science, 337(6096):8l6-2l (2012)). By transfecting a cell with elements including a Cas gene and specifically designed

CRISPRs, nucleic acid sequences can be cut and modified at any desired location. Methods of preparing compositions for use in genome editing using the CRISPR/Cas systems are described in detail in US Pub. No. 2016/0340661, US Pub. No. 2016/0340662, US Pub. No. 2016/0354487, US Pub. No. 2016/0355796, US Pub. No. 2016/0355797, and WO

2014/018423, which are specifically incorporated by reference herein in their entireties.

[0064] Zinc-finger nucleases (ZFNs) are artificial restriction enzymes generated by fusing a zinc finger DNA-binding domain to a DNA-cleavage domain. Zinc finger domains can be engineered to target specific desired DNA sequences and this enables zinc-finger nucleases to target unique sequences within complex genomes. By taking advantage of endogenous DNA repair machinery, these reagents can be used to precisely alter the genomes of higher organisms. The most common cleavage domain is the Type IIS enzyme Fokl. Fokl catalyzes double-stranded cleavage of DNA, at 9 nucleotides from its recognition site on one strand and 13 nucleotides from its recognition site on the other. See, for example, U.S. Pat. Nos. 5,356,802; 5,436,150 and 5,487,994; as well as Li et al. Proc., Natl. Acad. Sci. USA 89 (l992):4275-4279; Li et al. Proc. Natl. Acad. Sci. USA, 90:2764-2768 (1993); Kim et al.

Proc. Natl. Acad. Sci. USA. 91:883-887 (l994a); Kim et al. J. Biol. Chem. 269:31,978-31,982 (l994b), all of which are incorporated herein by reference. One or more of these enzymes (or enzymatically functional fragments thereof) can be used as a source of cleavage domains.

[0065] In some embodiments, a heme-enriched algae is created by genetically modifying a strain to modify the chlorophyll and/or heme pathways. Introduction of recombinant nucleic acids such as those that interfere with, inhibit or down-regulate expression of an endogenous gene (e.g., one or more of CHLD, CHLI1, CHLI2 or CHLH1) can alter the flux through the pathway. Such genetic modifications can include the integration of recombinant DNA in a regulatory region, exon or intron for an endogenous gene, as well as the gene silencing (e.g., introduction of antisense or siRNA for down regulating or silencing the expression of one or more endogenous genes). In some embodiments, expression of genes within the pathway can be upregulated such that the pathway produced more PPIX that can be converted to heme, or upregulates the expression or activity of ferrochelatase to produce more heme in the algae. Nucleic acids for modification of ferrochelatase can include the regulatory regions, such as those of SEQ ID NOs: 114, 115, exons, such as those of SEQ ID NOs: 116-122, and introns, such as those of SEQ ID NOs: 123-128. In some embodiments, a heme enriched algae may include an increased copy number of ferrocheletase or the provision of a construct to overexpress ferrocheletase (such as those provided by nucleic acid sequence SEQ ID NO: 7, and protein sequence SEQ ID NO: 8). In some embodiments, genetic modifications include modifications to or expression of one or more genes in the chloroplast. In some

embodiments, modifications are made to nuclear encoded genes or expression of such genes.

Algae Genus and Species for use in the Compositions and Methods

[0066] In the compositions and methods provided herein for producing heme and heme- containing compositions, algae strains that have a heme biosynthesis pathway are employed. In some embodiments, the algae strain for providing heme is a Chlorophyta (green algae). In some embodiments, the green algae is selected from the group consisting of Chlamydomonas, Dunaliella, Haematococcus, Chlorella, and Scenedesmaceae . In some embodiments, the Chlamydomonas is a Chlamydomonas reinhardtii. In varying embodiments, the green algae can be a Chlorophycean, a Chlamydomonas, C. reinhardtii, C. reinhardtii l37c, or a psbA deficient C. reinhardtii strain. In some embodiments, the selected host is Chlamydomonas reinhardtii, such as in Rasala and Mayfield, Bioeng Bugs. (2011) 2(l):50-4; Rasala, et al., Plant Biotechnol J. (2011) May 2, PMID 21535358; Coragliotti, et al., Mol Biotechnol. (2011) 48(l):60-75; Specht, et al., Biotechnol Lett. (2010) 32(10): 1373-83; Rasala, et al., Plant Biotechnol J. (2010) 8(6):7l9-33; Mulo, et al., Biochim Biophys Acta. (2011) May 2, PMID:2l565l60; and Bonente, et al, Photosynth Res . (2011) May 6, PMID:2l547493; US Pub. No. 2012/0309939; US Pub. No. 2010/0129394; and Intl. Pub. No. WO 2012/170125. All of the foregoing references are incorporated herein by reference in their entirety for all purposes.

[0067] In some embodiments, the algae strain for providing heme is a single-celled algae. Illustrative and additional microalgae species of interest include without limitation,

Achnanthes orientalis, Agmenellum, Amphiprora hyaline, Amphora coffeiformis, Amphora coffeiformis linea, Amphora coffeiformis punctata, Amphora coffeiformis taylori, Amphora coffeiformis tenuis, Amphora delicatissima, Amphora delicatissima capitata, Amphora sp., Anabaena, Ankistrodesmus, Ankistrodesmus falcatus, Boekelovia hooglandii, Borodinella sp., Botryococcus braunii, Botryococcus sudeticus, Carteria, Chaetoceros gracilis, Chaetoceros muelleri, Chaetoceros muelleri subsalsum, Chaetoceros sp., Chlamydomonas sp.,

Chlamydomonas reinhardtii, Chlorella anitrata, Chlorella Antarctica, Chlorella

aureoviridis, Chlorella Candida, Chlorella capsulate, Chlorella desiccate, Chlorella ellipsoidea, Chlorella emersonii, Chlorella fusca, Chlorella fusca var. vacuolata, Chlorella glucotropha, Chlorella infusionum, Chlorella infusionum var. actophila, Chlorella infusionum var. auxenophila, Chlorella kessleri, Chlorella lobophora (strain SAG 37.88), Chlorella luteoviridis, Chlorella luteoviridis var. aureoviridis , Chlorella luteoviridis var. lutescens, Chlorella miniata, Chlorella minutissima, Chlorella mutabilis, Chlorella nocturna, Chlorella parva, Chlorella photophila, Chlorella pringsheimii, Chlorella protothecoides, Chlorella protothecoides var. acidicola, Chlorella regularis, Chlorella regularis var.

minima, Chlorella regularis var. umbricata, Chlorella reisiglii, Chlorella saccharophila, Chlorella saccharophila var. ellipsoidea, Chlorella salina, Chlorella simplex, Chlorella sorokiniana, Chlorella sp., Chlorella sphaerica, Chlorella stigmatophora, Chlorella vanniellii, Chlorella vulgaris, Chlorella vulgaris, Chlorella vulgaris f. tertia, Chlorella vulgaris var. autotrophica, Chlorella vulgaris var. viridis, Chlorella vulgaris var. vulgaris, Chlorella vulgaris var. vulgaris f. tertia, Chlorella vulgaris var. vulgaris f. viridis, Chlorella xanthella, Chlorella zoflngiensis , Chlorella trebouxioides, Chlorella vulgaris, Chlorococcum infusionum, Chlorococcum sp. , Chlorogonium, Chroomonas sp., Chrysosphaera sp., Cricosphaera sp., Crypthecodinium cohnii, Cryptomonas sp., Cyclotella cryptica, Cyclotella meneghiniana, Cyclotella sp., Dunaliella sp., Dunaliella bardawil, Dunaliella bioculata, Dunaliella granulate, Dunaliella maritime, Dunaliella minuta, Dunaliella parva, Dunaliella peircei, Dunaliella primolecta, Dunaliella salina, Dunaliella terricola, Dunaliella tertiolecta, Dunaliella viridis, Dunaliella tertiolecta, Eremosphaera viridis, Eremosphaera sp.,

Ellipsoidon sp., Euglena, Franceia sp., Fragilaria crotonensis, Fragilaria sp., Gleocapsa sp., Gloeothamnion sp., Hymenomonas sp., Isochrysis aff. galbana, Isochrysis galbana,

Lepocinclis, Micractinium, Micractinium (UTEX LB 2614), Monoraphidium minutum, Monoraphidium sp., Nannochloris sp., Nannochloropsis salina, Nannochloropsis sp., Navicula acceptata, Navicula biskanterae, Navicula pseudotenelloides, Navicula pelliculosa, Navicula saprophila, Navicula sp., Nephrochloris sp., Nephroselmis sp., Nitschia communis, Nitzschia alexandrina, Nitzschia communis, Nitzschia dissipata, Nitzschia frustulum, Nitzschia hantzschiana, Nitzschia inconspicua, Nitzschia intermedia, Nitzschia microcephala, Nitzschia pusilla, Nitzschia pusilla elliptica, Nitzschia pusilla monoensis, Nitzschia quadrangular, Nitzschia sp., Ochromonas sp., Oocystis parva, Oocystis pusilla, Oocystis sp., Oscillatoria limnetica, Oscillatoria sp., Oscillatoria subbrevis, Pascheria acidophila,

Pavlova sp., Phagus, Phormidium, Platymonas sp., Pleurochrysis carterae, Pleurochrysis dentate, Pleurochrysis sp., Prototheca wickerhamii, Prototheca stagnora, Prototheca portoricensis , Prototheca moriformis, Prototheca zopfli, Pyramimonas sp., Pyrobotrys, Sarcinoid chrysophyte, Scenedesmus armatus, Schizochytrium, Spirogyra, Spirulina platensis, Stichococcus sp., Synechococcus sp., Tetraedron, Tetraselmis sp., Tetraselmis suecica, Thalassiosira weissflogii, and Viridiella fridericiana. In some embodiments, the algae is a Chlamydomonas species. In some embodiments, the algae is a Chlamydomonas reinhardtii. In some embodiments, the algae is a derivative of a green Chlamydomonas strain made by mutagenesis, by screening, by selection or by mating with another algae strain.

[0068] In some embodiments, the algae strain for use in the methods herein and for making heme-containing compositions is selected or identified based on one or more phenotypes and/or genotypes. In some embodiments, the algae strain for overproducing heme can be created through mating processes. In some embodiments, the algae strain for overproducing heme can be created through mutagenesis, such as ultra violet mutagenesis. In some embodiments, the algae strain for overproducing heme can be generated through chemical mutagenesis with a compound that results in DNA alterations.

[0069] Methods for selection of algae include, but are not limited to, genetic screening or phenotypic screening for deficiencies, mutations and changes in the chlorophyll biosynthesis pathway and/or chlorophyll accumulation, and by genetic screening or phenotypic screening for increased expression and/or accumulation of heme, heme biosynthesis intermediates and heme biosynthesis enzymes. In some embodiments, the algae strain for use in the methods herein and for making heme-containing compositions is selected or identified based on its spectral profile and/or its red or red-like color. In some embodiments, the algae for use in the methods herein and for making heme-containing compositions is selected or identified based on its growth rate in dark conditions. In some embodiments, the selection is based on growth rate in dark conditions and the appearance or enhancement of a red or red-like color when grown in dark conditions. In some embodiments, an algae strain is selected which is deficient in or reduced in the amount of carotenoids produced or accumulated.

[0070] In some embodiments, algae strains are mated to combine or enhance

characteristics that contribute to heme production, heme accumulation, reduction in chlorophyll and/or reduction in carotenoids. In some embodiments, an algae strain that has fast growth under dark conditions (e.g., faster than a wildtype strain) is mated with an algae strain that exhibits a red or red-like color. In some embodiments, an algae strain deficient for carotenoid production or accumulation is mated with an algae strain exhibiting a red or red- like color. [0071] In some embodiments, an algae strain is mutagenized and then a new strain is selected or identified that exhibits one or more characteristics of increased heme production, heme accumulation, reduction in chlorophyll and/or reduction in carotenoids. In some embodiments, an algae strain is generated by mutagenesis of a first starting strain and selection of a second strain that grows faster in the dark than the first starting strain. In some embodiments, an algae strain is generated by mutagenesis of a first starting strain and selection of a second strain that lacks one or more carotenoids. In some embodiments, the strain includes further modifications, such as a modification that decreases omega oils (e.g., omega-3 fatty acids) and/or a modification that allows the strain to grow on a particular carbon source such as glucose, dextrose, sucrose, etc.

[0072] In some embodiments, the algae is a Chlamydomonas species, such as

Chlamydomonas reinhardtii and the strain has a visible red or reddish-brown appearance. In some embodiments, the strain also exhibits growth on glucose. In some embodiments, the strain has a genetic modification in the chlorophyll synthetic pathway, such as in a nuclearly encoded subunit of Mg-chelatase , such as in a gene encoding CHLD, CHLI1, CHLI2 or CHLH1, or in an intron or regulatory region thereof, whereby the strain overexpresses or is enriched in heme content. In some embodiments, the strain is also enriched in PPIX content. In some embodiments, the strain is capable of growing to high culture density under fermentation conditions.

Culture methods for overproducing heme strains

[0073] Methods for growing algae in liquid media include a wide variety of options including ponds, aqueducts, small scale laboratory systems, and closed and partially closed bioreactor systems. Algae can also be grown directly in water, for example, in an ocean, sea, lake, river, reservoir, etc.

[0074] In some embodiments, the heme overproducing algae useful in the methods and compositions provided herein are grown in a controlled culture system, such as a small scale laboratory systems, large scale systems and closed systems and partially closed bioreactor systems. Small scale laboratory systems refer to cultures in volumes of less than about 6 liters, and can range from about 1 milliliter or less up to about 6 liters. Large scale cultures refer to growth of cultures in volumes of greater than about 6 liters, and can range from about 6 liters to about 200 liters, and even larger scale systems covering 5 to 2500 square meters in area, or greater. Large scale culture systems can include liquid culture systems from about 10,000 to about 20,000 liters and up to about 1,000,000 liters.

[0075] The culture systems for use with the methods for producing the compositions herein include closed structures such as bioreactors, where the environment is under stricter control than in open systems or semi-closed systems. A photobioreactor is a bioreactor which incorporates some type of light source to provide photonic energy input into the reactor. The term bioreactor can refer to a system closed to the environment and having no direct exchange of gases and contaminants with the environment. A bioreactor can be described as an enclosed, and in the case of a photobioreactor, illuminated, culture vessel designed for controlled biomass production of liquid cell suspension cultures.

[0076] In some embodiments, the algae used in the methods and for the compositions provided herein are grown in fermentation vessels. In some embodiments, the vessel is a stainless steel fermentation vessel. In some embodiments, the algae are grown in

heterotrophic conditions whereby one or more carbon sources is provided to the culture. In some embodiments, the algae are grown in aerobic and heterotrophic conditions. In some embodiments, the algae are grown to a density greater than or about lOg/L, about 20 g/L, about 30 g/L, about 40g/L, about 50g/L, about 75 g/L, about lOOg/L, about 125 g/L, or about l50g/L.

[0077] In some embodiments, the algae are inoculated from a seed tank to a starting density of greater than about O.lg/L, about LOg/L, about 5.0g/L, about lO.Og/L, about 20.0g/L, about 50g/L, about 80g/L, or about lOOg/L. Once inoculated, the algae are grown heterotrophically using an aerobic fermentation process. During this process, the algae are fed nutrients to maintain their growth. In some embodiments, these nutrients include a reduced carbon source. Exemplary aerobic fermentation process and/or reduced carbon sources include, but are not limited to, acetate, glucose, sucrose, fructose, glycerol and other types of sugars ( e.g ., dextrose, maltose, galactose, sucrose, ribose, etc.). In some

embodiments, the algae culture is supplemented with iron.

[0078] In some embodiments, the algae are cultured under dark conditions. Preferably, the dark condition has a brightness of less than 1000 lux, less than 750 lux, less than 500 lux, less than 400 lux, less than 300 lux, less than 200 lux, less than 100 lux. In some embodiments, the algae cultured under dark conditions lack or are reduced in chlorophyll production at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, or at least 80% compared to the algae cultured under dark conditions. In some embodiments, the algae grown under dark conditions are supplemented with one or more nutrients. In some embodiments, the algae grown under dark conditions are grown in the presence of a reduced carbon source, such as acetate, glucose, sucrose, fructose, glycerol or other types of sugars (e.g., dextrose, maltose, galactose, sucrose, ribose, etc.). In some embodiments, the algae grown under dark conditions are grown in the presence of iron or otherwise supplemented with iron.

[0079] In some embodiments, the heme-enriched strains herein are grown in dark or limited light conditions such that the pathway flux to biliverdin IX and photochromobilin are decreased, and the amount of heme in such strains is increased. In some embodiments, the heme-enriched strains herein are grown in dark or limited light condition and utilize a carbon source such as glucose.

Edible Food Products and Ingredients

[0080] Provided herein are edible products for human and animal consumption that contain heme from algae. In some embodiments, the edible product is a beef-like product, a fish-like product or a meat replica. In some embodiments, the edible product contains whole cell algae, where the algae provides heme to the composition. In some embodiments, the heme is imparted to the edible product by a whole cell algae component where the algae overproduce heme. In some embodiments, the heme is imparted to the edible product by an algae having a heme content greater than the chlorophyll content of the algae. In some embodiments, the heme is imparted to the edible product by an algae having a protoporphyrin content greater than chlorophyll content by at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, or at least 50%.

[0081] In some embodiments, the edible product is a beef-like product, a fish-like product or a meat replica and the heme is provided by fractionated algae. For example, whole cell alga producing or overproducing heme can be subjected to fractionation methods to separate some or a substantial amount of biomass from the heme-containing fraction. The fractionation may remove one or more components of the algae biomass while leaving other components such as omega-3 fatty acids, fats, protein, vitamin A, beta-carotene or any combination thereof associated with the heme-containing fraction. In some embodiments, the heme can be separated from one or more of the omega-3 fatty acids, saturated fats, protein, vitamin A, and/or beta-carotene of the algae. Extraction with solvents and buffers or a combination thereof can be used to provide a heme-enriched fraction. For example, an alga biomass or a fractions thereof can be enriched for heme through hexane extraction.

[0082] In some embodiments, the biomass is fractionated or otherwise treated to separate heme content and optionally, PPIX. Such fractionation can include separation of PPIX from heme. For example, heme-binding proteins and heme associated with proteins can be separated from PPIX which is not a protein-conjugated or protein-associated compound.

Both free heme and protein-associated heme can be separated from PPIX based on heme’s association with iron. PPIX does not contain an iron moiety and as such, this feature can be used to separate PPIX from a heme-containing fraction. In some embodiments, an algae biomass herein is fractionated or otherwise treated such that the heme is separated from other components, including PPIX.

[0083] In some embodiments, the heme-containing fraction has a heme content greater than the chlorophyll content of the fraction by at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, or at least 50%. In some embodiments, the heme-containing fraction has a protoporphyrin IX content greater than chlorophyll content of the fraction by at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, or at least 50%. In some embodiments, the heme-containing fraction contains no chlorophyll or substantially no chlorophyll. In some embodiments, the heme-containing fraction has no chlorophyll or substantially no chlorophyll and has about 4.5% protoporphyrin IX content (on a weight per total weight basis, e.g., 45 mg protoporphyrin IX in a 1 gram sample). In some embodiments, the heme- containing fraction has no chlorophyll or substantially no chlorophyll and has about 0.5% heme content (on a weight per total weight basis, e.g., 5 mg heme in a 1 gram sample). In some embodiments, the heme-containing fraction has no chlorophyll or substantially no chlorophyll and has about 4.5% protoporphyrin IX content and has about 0.5% heme content (on a weight per total weight basis).

[0084] In some embodiments, a whole algae preparation used in the preparation of an edible composition has a heme content greater than the chlorophyll content of the fraction. In some embodiments, the whole algae preparation has a protoporphyrin IX content greater than chlorophyll content of the fraction. In some embodiments, the whole algae preparation contains no chlorophyll or substantially no chlorophyll. In some embodiments, the whole algae preparation has no chlorophyll or substantially no chlorophyll and has about 4.5% protoporphyrin IX content (on a weight per total weight basis, e.g., 45 mg protoporphyrin IX in a 1 gram sample). In some embodiments, the whole algae preparation has no chlorophyll or substantially no chlorophyll and has about 0.5% heme content (on a weight per total weight basis, e.g., 5 mg heme in a 1 gram sample). In some embodiments, the whole algae preparation has no chlorophyll or substantially no chlorophyll and has about 4.5%

protoporphyrin IX content and has about 0.5% heme content (on a weight per total weight basis).

[0085] In some embodiments, the whole algae preparation or fractionated algae preparation has no chlorophyll or substantially no chlorophyll and is made from an algae strain that does not make or accumulate chlorophyll. In some embodiments, the whole algae preparation or fractionated algae preparation has no chlorophyll or substantially no chlorophyll and is made from an algae strain that has one or more mutations in the chlorophyll synthesis pathway and/or has one or more mutations in the pathways that impact the accumulation or turnover of chlorophyll, for example, having a modification in one or more subunits of magnese chelatase such as a modification in one or more of CHLD, CHLI1, CHLI2 or CHLH1.

[0086] In some embodiments, the whole algae preparation or fractionated algae preparation contains heme at about 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2.0%, 2.5% or more than 2.5% on a weight per total weight basis. In some embodiments, the whole algae preparation or fractionated algae preparation contains protoporphyrin IX at about 0.5%, 1.0%, 1.5%,

2.0%, 2.5%, 3.0%, 3.5%, 4.5%, 5.0%, 5.5%, 6.0%, 6.5%, 7.0%, 7.5%, 8.0%, 8.5%, 9.0%, 9.5%, 10.0% or more than 10% on a weight per total weight basis. In some embodiments, the heme in the whole algae preparation or fractionated algae preparation is free heme. In some embodiments, the heme in the whole algae preparation or fractionated algae preparation is complexed with one or more proteins, for example complexed to one or more truncated hemoglobins. In some embodiments, the heme in the whole algae preparation or fractionated algae preparation is a mixture of free heme and heme complexed with protein.

[0087] In some embodiments, the whole cell or fractionated algae provides protein to the edible composition as well as providing heme. In some embodiments, the algae provides at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9% or 10% of the protein to the edible composition. In some embodiments, the algae provides greater than about 5%, 10%, 15%,

20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%,

99% or 100% of the protein in the edible product. In some embodiments, the whole cell or fractionated algae provides protein to the edible composition and the edible composition also contains protein from one or more additional sources, such as a plant-based source. In some embodiments, an alga fraction is enriched for protein as compared to the starting biomass hexane extraction or an equivalent solvent can be used to enrich the protein content of the fraction. In some embodiments, carbohydrates and/or fatty acids are removed or reduced in amount through such extraction(s), while enriching for protein and/or enriching for heme.

[0088] In some embodiments, the whole cell or fractionated algae provides omega-3 fatty acids to the edible composition as well as providing heme. In some embodiments, the algae provides a daily recommended dosage of omega-3 fatty acids or a portion thereof to the edible product. For example, the whole cell or fractionated algae provides at least about 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg, 95 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, or 500 mg of omega-3 fatty acids to the edible composition.

[0089] In some embodiments, omega oils such as omega-3 fatty acids are removed from the alga biomass or a fractionated alga sample. Such oil removal can modify the aroma and taste of the alga biomass or faction, such as by decreasing or removing a“fishy” aroma or taste that can be present in an alga-derived product. In some embodiments, hexane or a similar solvent such as isohexane, heptane, butane or other alcohol, is used in the preparation of the alga biomass or fractionation to modify the aroma and taste. In some cases, hexane or similar solvent extraction removes or decreases the amount of oils, as well as enriches for heme and/or enriches for protein in the resulting product.

[0090] In some embodiments, algae biomass or fractionate algae are made using a strain deficient in one or more omega oils. Such strains can be combined with a heme-enriched strain, such as through mating to produce a heme-enriched strain that produces less omega oils.

[0091] In some embodiments, the whole cell or fractionated algae provides vitamin A to the edible composition as well as providing heme. In some embodiments, the algae provides a daily recommended dosage of vitamin A or a portion thereof to the edible product. For example, the whole cell or fractionated algae provides at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or 100% of the daily recommended dosage of vitamin A or at least about 20 pg, 50 pg, 100 pg, 200 pg, 300 pg, 400 pg, 500 pg, 600 pg, 700 pg, 800 pg, 900 pg or 1000 pg of retinol activity equivalents (RAE) for vitamin A. In some embodiments, the whole cell or fractionated algae provides no more than about 2,000 pg, 2,500 pg or 3,000 pg of retinol activity equivalents (RAE) for vitamin A.

[0092] In some embodiments, the whole cell or fractionated algae provides beta-carotene to the edible composition as well as providing heme. In some embodiments, the algae provides a daily recommended dosage of beta-carotene or a portion thereof to the edible product. For example, the whole cell or fractionated algae provides at least about 5%, 10%,

15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%.

95%, 99% or 100% of the daily recommended dosage of beta-carotene. In some

embodiments, the algae provides about 0.25 mg, 0.5 mg, 1 mg, 1.5 mg, 2 mg, 2.5 mg, 3 mg, 4 mg, 5, mg, 6 mg, 9 mg, 10 mg, 12 mg, or 15 mg of beta-carotene.

[0093] In some embodiments, the whole cell or fractionated algae that provides heme contains saturated fat. In some embodiments, the algae provides less than daily

recommended limit for saturated fat or a portion thereof to the edible product. For example, the whole cell or fractionated algae provides no more than about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the daily recommended dosage of saturated fat. In some embodiments, the algae provides no more than 0.01%, 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.2%, 1.5%, 2%, 5% or 10% of total saturated fat present in the edible composition or in the finished product made from the edible composition.

[0094] In some embodiments herein, the heme-containing whole algae or algae fraction is used to create an edible composition that is then used as an ingredient in a finished product. The ingredient may provide heme as well as omega-3 fatty acids, fats, protein, vitamin A, beta-carotene or any combination thereof to the ingredient. Such ingredient may be a colorant, texturant, binder, nutrient source, taste or flavor enhancer, or a filler. [0095] In some embodiments, the heme-containing whole algae or algae fraction is used to create an edible composition that is a finished product. For example, the finished product may be a meat-like product such as a burger, a patty, a cake, a ground“meat,” a sausage, a kebab, a steak, cubed“meat,” a“meatball,” a filet, a drumstick, a“chicken finger,” or a “chicken nugget.” The finished product may be a meat-like product made to resemble beef, chicken, pork, wild game, turkey or other consumable meat product. The finished product may be a fish-like product made to resemble a fish filet, a fish patty or cake, a fish ball, a fish salad, ground fish, a fish nugget, a fish burger or the like, such as a tuna product, a spicy tuna product or a salmon product.

[0096] The whole algae or algae fraction may provide omega-3 fatty acids, saturated fats, protein, vitamin A, beta-carotene or any combination thereof to the finished product. In some embodiments, the whole algae or algae fraction can be reduced in omega oils and used for the finished product. Meat-like products can be made with a whole algae or algae fraction from a heme-enriched algae that is as described herein, by processing or by strain type, reduced in the amount of omega oils.

[0097] In some embodiments, the finished product comprising the whole algae or algae fraction is a cooked product. In some embodiments, the finished product comprising the whole algae or algae fraction is a uncooked product or raw product. In some embodiments, the finished product comprising the whole algae or algae fraction is a partially-cooked product.

Heme-containing preparations and products

[0098] Algae strains and cultures overproducing heme such as described herein can be used in various forms and preparations. In some embodiments, a heme-containing composition is prepared from an algae culture overproducing heme, where the composition is red or red-like in color.

[0099] In some embodiments, the heme-containing composition is prepared from a biomass isolated from cultured algae. In some embodiments, the biomass is further fractionated to remove one or more components. In some embodiments, the biomass is fractionated to remove starch. In some embodiments, the biomass is fractionated to remove protein. In some embodiments, the biomass is fractionated or otherwise treated to remove carotenoids. In some embodiments, the biomass is fractionated or otherwise treated to enrich for certain components. In some embodiments, the fractionated or treated biomass is enriched in heme. In some embodiments, the fractionated or treated biomass is enriched in protein or in protein and heme. In some embodiments, the fractionation or treatment enhances the red or red-like color of the preparation. The fractionated or treated biomass can be enriched for protein content such that the composition is about 10% protein, greater than about 10% protein, or greater than about 20%, about 30%, about 40%, or about 50% protein.

[0100] In some embodiments, the heme-containing composition is a heme-containing liquid prepared from the culture media of the cultured algae. In some embodiments, the heme-containing composition is prepared from heme found extracellularly in the algae culture. In some embodiments, the algae culture is lysed or otherwise treated to release heme from the cells. In some embodiments, the heme-containing liquid is further fractionated to remove one or more components. In some embodiments, the heme-containing liquid is fractionated to remove starch. In some embodiments, the heme-containing liquid is fractionated to remove protein. In some embodiments, the heme-containing liquid is fractionated or otherwise treated to remove carotenoids. In some embodiments, the heme- containing liquid is fractionated or otherwise treated to enrich for certain components. In some embodiments, the fractionated or treated heme-containing liquid is enriched in heme.

In some embodiments, the fractionation or treatment enhances the red or red-like color of the preparation.

[0101] The heme-containing compositions, including biomass, liquid and fractionated preparations can be further processed. Such processing can include concentrating, drying, lyophilizing, and freezing. In various embodiments, the heme-containing compositions can be combined with additional components and ingredients. In some embodiments, the heme- containing composition is combined with additional ingredients to create an edible product.

In some embodiments, the heme-containing composition confers a red or red-like color to the edible product. In some embodiments, the heme-containing composition confers a meat-like characteristic such as a meat-like taste, meat-like flavor aroma and/or texture to the edible product. In some embodiments, the heme-containing composition provides the appearance of blood to an edible product, such as to a meat replica, a beef-like product, a chicken-like product or the like. Alternatively, at least one of the features of meat or meat-like flavor or aroma, a meat or meat-like texture, a blood-like appearance, a meat or meat-like color are derived from the algae preparation. [0102] In some embodiments, heme-containing compositions are combined with additional ingredients to create a meat-like product. Such meat-like products can include clean meat or cultured meat (made from animal cells grown in the laboratory or otherwise outside of an animal), plant-based and non-animal based meats (made from plant ingredients and/or ingredients not from animal sources). In some embodiments, a heme-containing composition made from an over-producing algae is combined with additional ingredients to create a meat-like product whereby the addition of the heme-containing composition confers a red or red-like color, a meat-like aroma, a meat-like taste and/or a meat-like texture to the meat-like product. In some embodiments, the meat-like features conferred by the heme- containing composition are conferred to the raw or uncooked product. In some embodiments, the meat-like features conferred by the heme-containing composition is conferred to the cooked product.

[0103] In some embodiments, whole algae or fractionated algae is combined with an additional protein source in an edible composition. For example, the protein source is wheat protein, such as wheat protein textured wheat protein, pea protein, textured pea protein, soy protein, textured soy protein, potato protein, whey protein, yeast extract, or other plant-based protein source or any combination thereof. In some embodiments, whole algae or fractionated algae is combined with an oil or source of fat in an edible composition. For example, the oil or fat source is coconut oil, canola oil, sunflower oil, safflower oil, com oil, olive oil, avocado oil, nut oil or other plant-based oil or fat source or any combination thereof. In some embodiments, whole algae or fractionated algae is combined with a starch or other carbohydrate source such as from potato, chickpea, wheat, soy, beans, com or other plant-based starch or carbohydrate or any combination thereof. In some embodiments, whole algae or fractionated algae is combined with a thickener in an edible composition. For example, starches as arrowroot, cornstarch, katakuri starch, potato starch, sago, tapioca and their starch derivatives may be used as a thickener; microbial and vegetable gums used as food thickeners include alginin, guar gum, locust bean gum, konjac and xanthan gum; and proteins such as collagen and egg whites may be used as thickeners; and sugar polymers for use as thickeners include agar, methylcellulose, carboxymethyl cellulose, pectin and carrageenan. In some embodiments, whole algae or an algae fraction may be combined with vitamins and minerals in an edible composition, such as vitamin E, vitamin C, thiamine (vitamin Bl), zinc, niacin, vitamin B6, riboflavin (vitamin B2), and vitamin B12. [0104] In some embodiments, whole algae or an algae fraction may be combined with additional ingredients such that the edible composition and/or finished product is vegetarian, vegan or gluten-free and therefore may conform to the dietary guidelines of Jewish kosher practitioners, and halal practitioners. Thus, in some embodiments, the edible composition and/or finished product may be suitable for consumption by vegetarians, vegans, gluten-free populations, Jewish kosher practitioners, and halal practitioners. In some embodiments, whole algae or an algae fraction may be combined with additional ingredients such that the edible composition and/or finished product is GMO-free and/or does not contain any ingredients derived from genetically engineered organisms or cells.

EXEMPLARY NUMBERED EMBODIMENTS

[0105] The following embodiments recite non-limiting permutations of combinations of features disclosed herein. Other permutations of combinations of features are also contemplated. In particular, each of these numbered embodiments is contemplated as depending from or relating to every previous or subsequent numbered embodiment, independent of their order as listed.

[0106] Embodiment 1. An engineered algae having a genetic modifications, where the genetic modification results in an accumulation of heme in the algae as compared to an algae lacking the genetic modification. 2. The engineered algae of embodiment 1, wherein the engineered algae has reduced or absence of chlorophyll production. 3. The engineered algae of embodiment 1 or embodiment 2, wherein the algae has red or red-like color. 4. The engineered algae according to any of embodiments 1-3, wherein the algae is capable of growth on glucose as the sole carbon source. 5. The engineered algae according to any of embodiments 1-4, wherein the genetic modification comprises a genetic alteration to chlorophyll synthesis pathway, protoporphyrinogen IX synthesis pathway or heme synthesis pathway. 6. The engineered algae according to any of embodiments 1-5, wherein the genetic modification is associated with a deficiency in the expression of magnesium chelatase. 7. The engineered algae according to any of embodiments 1-6, wherein the genetic modification comprises an alteration in one or more of CHLD, CHLI1, CHLI2 or CHLH1. 8. The engineered algae of embodiment 7, wherein the genetic modification comprises an alteration in an upstream regulatory region, a downstream regulatory region, an exon, an intron or any combination thereof. 9. The engineered algae according to any of embodiments 5-8, wherein the genetic modification comprises an insertion, a deletion, a point mutation, an inversion, a duplication, a frameshift or any combination thereof. 10. The engineered algae according to any of embodiments 1-9, wherein the engineered algae has a heme content greater than the chlorophyll content. 11. The engineered algae according to any of embodiments 1-10, wherein the engineered algae has a protoporphyrin IX content greater than the chlorophyll content. 12. The engineered algae according to any of embodiments 1-11, wherein the engineered algae has reduced production of one or more fatty acids. 13. The engineered algae according to any of embodiments 1-12, wherein the engineered algae further comprises a genetic modification that reduces or eliminates the expression of light independent protochlorophyllide oxidoreductase. 14. The engineered algae of embodiment 13, wherein the genetic modification comprises a mutation or deletion in one or more of ChlB, ChlL or ChlN. 15. The engineered algae according to any of embodiments 1-14, wherein the engineered algae has upregulated expression of ferrocheletase. 16. The engineered algae according to any of embodiments 1-15, wherein the engineered algae has upregulated expression of protoporphyrinogen IX oxidase. 17. The engineered algae according to any of embodiments 1-16, wherein the algae contain a recombinant or heterologous nucleic acid. 18. The engineered algae according to any of embodiments 1-17, wherein the engineered algae comprises a Chlamydomonas sp. 19. The engineered algae of embodiment 18, wherein the Chlamydomonas sp. is Chlamydomonas reinhardtii.

[0107] Embodiment 20. An edible composition comprising an algae preparation, wherein the algae preparation comprises an engineered algae of any of embodiments 1-19 or a portion thereof. 21. The edible composition of embodiment 20, wherein the edible composition comprises heme derived from the engineered algae. 22. The edible composition of embodiment 20, wherein the algae preparation comprises algae cells. 23. The edible composition of embodiment 20, wherein the algae preparation is a fractionated algae preparation. 24. The edible composition according to any of embodiments 20-23, wherein the algae preparation is red or red-like in color. 25. The edible composition according to any of embodiments 20-24, wherein the edible composition has a red or red-like color derived from the algae preparation. 26. The edible composition according to any of embodiments 20-25, wherein the algae preparation confers a meat or meat-like flavor to the edible composition.

27. The edible composition according to any of embodiments 20-26, wherein the edible composition has a meat or meat-like texture derived from the algae preparation. 28. The edible composition according to embodiment 27, wherein the meat or meat-like texture comprises a beef or beef-like texture, a fish or fish-like texture, a chicken or chicken-like texture, a pork or pork-like texture or a texture of a meat replica. 29. The edible composition according to any of embodiments 20-28, wherein the edible composition is a finished product selected from the group consisting of a beef-like food product, a fish-like product, a chicken- like product, a pork-like product and a meat replica. 30. The edible composition according to any of embodiments 20-29, wherein the edible composition is vegan, vegetarian or gluten- free. 31. The edible composition according to any of embodiments 20-30, wherein the edible composition has an appearance of blood derived from the algae preparation. 32. The edible composition according to any of embodiments 20-31, wherein the algae preparation has a heme content greater than the chlorophyll content. 33. The edible composition according to any of embodiments 20-32, wherein the algae preparation has a protoporphyrin IX content greater than the chlorophyll content. 34. The edible composition according to any of embodiments 20-33, wherein the algae preparation provides at least 5%, 10%, 15%, 20%,

25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or 100% of the total protein content to the edible composition. 35. The edible composition according to any of embodiments 20-34, wherein the algae preparation provides vitamin A, beta carotene or a combination thereof to the composition. 36. The edible composition of embodiment 35, wherein the vitamin A, the beta carotene or the combination thereof is at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%,

75%, 80%, 85%, 90%, 95%, 99% or 100% of the daily recommended requirement. 37. The edible composition according to any of embodiments 20-36, wherein the algae preparation provides less than about 0.01%, 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%,

0.9%, 1.0%, 1.2%, 1.5%, 2%, 5% or 10% of total saturated fat present in the edible composition. 38. The edible composition according to any of embodiments 20-37, wherein the algae preparation provides less than about 0.01%, 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%,

0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.2%, 1.5%, 2%, 5% or 10% of total saturated fat present in a finished product comprising the edible composition. 39. The edible composition according to any of embodiments 20-38, wherein the algae preparation provides at least about 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg, 95 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg or 500 mg of omega-3 fatty acids to the edible composition. 40. The edible composition according to any of embodiments 20-39, wherein the algae preparation has reduced fatty acid content. 41. The edible composition according to any of embodiments 20-40, wherein the edible product is combined with a protein source, a fat source, a carbohydrate, a starch, a thickener, a vitamin, a mineral, or any combination thereof. 42. The edible composition of embodiment 41, wherein the protein source is selected from the group consisting of textured wheat protein, textured soy protein and textured pea protein, fungal protein or algal protein. 43. The edible composition of embodiment 41, wherein the fat source comprises at least one of refined coconut oil or sunflower oil. 44. The edible composition of any of embodiments 41-43, further comprising at least one of potato starch, methylcellulose, water, and a flavor, wherein the flavor is selected at least one of yeast extract, garlic powder, onion powder, and salt. 45. The edible composition of any of embodiments 41-44, wherein the edible product is an ingredient for a burger, a sausage, a kebab, a filet, a fish-altemative, a ground meat-like product or a meatball. 46. The edible composition of embodiment 45, wherein the burger comprises about 5% of the algae preparation, about 20% textured soy protein and about 20% refined coconut oil. 47. The edible composition of embodiment 46, further comprising about 3% sunflower oil, about 2% potato starch, about 1% methylcellulose, about 45% water and about 4-9% flavors. 48. The edible composition of embodiment 46, further comprising about 0.5% Kojac gum, about 0.5% Xanthan gum, about 45% water and about 4-9% flavors. 49. The edible composition of embodiment 45, wherein the fish-altemative comprises 20% textured soy protein, about 5 % of algae preparation, about 65% water and about 10% flavors. 50. The edible composition according to any of embodiments 20-49, wherein the edible composition is free of animal proteins. 51. The edible composition according to any of embodiments 20-50, wherein the algae preparation comprises an algae having an increase in protoporphyrinogen IX synthesis or accumulation. 52. The edible composition according to any of embodiments 20-51, wherein the algae preparation comprises an algae that exhibits a red or red-like color when grown in the dark conditions. 53. The edible composition according to any of embodiments 20-52, wherein the algae comprised in the algae preparation are recombinant or genetically modified algae. 54. The edible composition according to any of embodiments 20-53, wherein the algae preparation comprises a Chlamydomonas sp. 55. The edible composition of embodiment 54, wherein the Chlamydomonas sp. is Chlamydomonas reinhardtii.

[0108] Embodiment 56. A method for the production of an edible composition comprising: (a) culturing an engineered algae according to any of embodiments 1-19 in a condition where the engineered algae exhibits a red or red-like color and wherein the engineered algae produces heme; (b) collecting the cultured engineered algae to produce an algae preparation; and (c) combining the algae preparation with at least one edible ingredient to produce an edible composition. 57. The method of embodiment 56, wherein the condition comprises a fermentation condition. 58. The method according to any of embodiments 56-57, wherein the condition comprises acetate as a reduced carbon source for growth of the engineered algae. 59. The method according to any of embodiments 56-58, wherein the condition comprises sugar as a reduced carbon source for growth of the engineered algae. 60. The method according to any of embodiments 56-59, wherein the condition comprises dark or limited light conditions. 61. The method according to any of embodiments 56-60, wherein the method further comprises fractionating the cultured algae to produce the algae preparation. 62. The method according to any of embodiments 56-61, wherein the algae preparation has a heme content that is greater than the chlorophyll content. 63. The method according to any of embodiments 56-62, wherein the algae preparation has a protoporphyrin IX content that is greater than the chlorophyll content. 64. The method according to any of embodiments 56-63, wherein the condition further comprises iron supplements. 65. The method according to any of embodiments 56-64, wherein the engineered algae is a

Chlamydomonas sp. 66. The method of embodiment 65, wherein the engineered algae is a Chlamydomonas reinhardtii. 67. The method according to any of embodiments 56-66, wherein the edible composition has at least one of the features selected from the group consisting of a meat or meat-like flavor, a meat or meat-like texture, a blood-like appearance and a meat or meat-like color, where the at least one of the features is derived from the algae preparation. 68. The method according to any of embodiments 56-67, wherein the method further comprises producing a finished product comprising the edible composition and wherein the finished product is a beef-like food product, a fish-like product, a chicken-like product, a pork-like product or a meat replica. 69. The method according to any of embodiments 56-68, wherein the edible composition is free of animal proteins. 70. The method according to any of embodiments 56-69, wherein the algae preparation is fractionated to remove one or more of starch, protein, PPIX, fatty acids and chlorophyll.

[0109] Embodiment 71. A method of making an engineered algae enriched in heme content, comprising: (a) subjecting an algae strain to a process that produces genetic modification to create a first algae population; and (b) from the first algae population, selecting a second algae population that is enriched in heme content, and optionally, PPIX content. 72. The method according to embodiment 71, wherein the process comprises at least one of a random UV mutagenesis, a random chemical mutagenesis, a recombinant genetic engineering, a gene editing, or a gene silencing. 73. The method according to embodiment 71 or embodiment 72, further comprising culturing the first algae population in a fermentation condition. 74. The method according to embodiment 73, wherein the fermentation condition comprises a media having sugar as a sole carbon source. 75. The method according to embodiment 74, wherein the sugar is selected from glucose, dextrose, fructose, maltose, galactose, sucrose, and ribose. 76. The method according to any of embodiments 73-75, wherein the fermentation condition comprises a brightness of less than 500 lux. 77. The method of any of embodiments 73-76, wherein the selecting the second algae population comprises sorting or identifying algae cells having a red or red-like color.

78. The method of any of embodiments 73-77, wherein the selecting is performed by FACS.

79. The method according to any of embodiments 73-78, the second algae population is selected with its capability to grow in the fermentation condition.

EXAMPLES

Example 1: Mutagenesis of algae and selection of strains

[0110] A wildtype strain of algae ( Chlamydomonas sp.) was subjected to UV irradiation with an excitation wavelength of 420nm and an emission of 635nm. Strains were first selected for their ability to grow on alternatives carbon sources such as glucose. One of these selected strains was further mutagemzed using similar conditions to select and/or identify for red-colored strains using fluorescence screening (e.g., Fluorescence-activated ceil sorting (FACS)) or magnetic or bead-based cell sorting. These selections are illustrated in FIG. 2 and as further detailed below.

[0111] Strains of algae ( Chlamydomonas reinhardtii) overexpressing heme were identified by their inability to produce chlorophyll. Additionally, these strains exhibited red, brown, orange or some variation of the listed color. The identified strains exhibit light sensitivity and cannot be grown in direct light greater than 10 mE m ² s ³ for extended periods of time.

[0112] To generate strains of algae overexpressing heme, green parental strains of Chlamydomonas reinhardtii were placed in a UV -light cross linker and exposed to 25 - 300mJ/cm ² of UV-light to induce random mutations. Following the exposure to UV-light strains were recovered on agar plates and placed into the dark. Once recovered, the strains were pulled into a flask with growth media and grown placed in a shaker in the dark to limit their potential for exposure to light which could cause many of the heme rich strains to be lost. Flask for cultured for a week in the dark and then applied to a flow cytometer. Cells were excited with a 420nm light and excitation was measured at 595±l5nm and 635±l5nm. Cells that had a high excitation signal at 595±l5nm were avoided as this the fluorescent signal for Mg-protoporphyrin, a precursor to the formation of chlorophyll. Cells that had a high fluorescent excitation signal at 635±l5nm were sorted into a pulled population as this fluorescent signal is indicative of high protoporphyrin IX. Once pulled, cells were spread on a plate an individual colonies grown and their individual fluorescent characteristics determined by a 96-well plate reader. This process resulted in the identification of 50 strains that had elevated levels of protoporphyrin IX and heme.

[0113] One of these red strains was subjected to genomic sequencing at the loci involved in chlorophyll and heme biosynthesis. Sequencing indicated that the genetic modification occurred in the CHLH locus. The sequence of CHLH of the red strain is provided in SEQ ID NO: 129 (nucleotide sequence) and SEQ ID NO: 152 (amino acid sequence). The modification deletes a single base pair in CHLH as compared to a green strain, causing a frameshift in the CHLH open reading frame and/or generate a stop codon such that the protein is translated into a truncated form. The sequence comparison is shown in FIG. 9 (upper sequence (Seq l) is a partial nucleic acid sequence (residues 1621-1679 of SEQ ID NO: 27) and a partial amino acid sequence (residues 451-460 of SEQ ID NO: 28) of CHLH gene of green algae, and lower sequence (Seq_2) is a partial nucleic acid sequence (residues 1621-1680 of SEQ ID NO: 129) and partial amino acid sequence (residues 451-460 of SEQ ID NO: 152) of CHLH gene of red algae has a mutation (asterisk)). The nucleic acid sequences of additional genes that may be altered in such algae strains are provided herein.

Example 1A: Identification of heme rich Chlamydomonas sp. that grow on sugar as their sole reduced carbon source

[0114] The use of sugar as a carbon source versus acetate has an economic benefit to the cost of production Chlamydomonas algae. To date, no strains of Chlamydomonas reinhardtii have been identified that grow on sugar as a carbon source. Typically, as shown in FIG. 3, Chlamydomonas reinhardtii requires acetate or sunlight and carbon dioxide to grow. Strains of algae from the wild or various culture collection centers were plated on agar growth media with dextrose added at 25g/L. The plates were then placed in the dark to ensure that photosynthesis could not occur. Cultures were allowed to grow for 2 weeks. At the end of two weeks cultures were studied for their ability to grow in conditions devoid of light. Strains that were capable of growing in the dark with dextrose as their primary carbon source were then placed into shake flasks with growth medium and dextrose at 25g/L as the primary carbon source and growth for a week in the dark. Culture density and sugar concentration in the media was monitored daily to determine if dextrose was being metabolized by the strains.

[0115] Following their identification, Chlamydomonas sp. strains that grew on dextrose as a carbon source were mutagenized using a UV-crosslinker. Cultures were exposed to 25 - 300mJ/cm ² of UV-light to induce mutations. Following the exposure to UV -light strains were recovered on agar plates and placed into the dark. Once recovered, the strains were pulled into a flask with growth media and grown placed in a shaker in the dark to limit their potential for exposure to light which could cause many of the heme rich strains to be lost. Flask for cultured for a week in the dark and then applied to a flow cytometer. Cells were excited with a 420nm light and excitation was measured at 595±l5nm and 635±l5nm. Cells that had a high excitation signal at 595±l5nm were avoided as this the fluorescent signal for Mg-protoporphyrin, a precursor to the formation of chlorophyll. Cells that had a high fluorescent excitation signal at 635±l5nm were sorted into a pulled population as this fluorescent signal is indicative of high protoporphyrin IX. Once pulled, cells were spread on a plate an individual colonies grown and their individual fluorescent characteristics determined by a 96-well plate reader. This process resulted in the identification of 20 strains that had elevated levels of protoporphyrin IX and heme and that were still able to grow on dextrose.

[0116] Tables 1-5 show characteristic analysis of one exemplary, identified red heme algae (Strain number: TAI114, Species name: Chlamydomonas reinhardtii).

TABLE 1: MICROBIAL ANALYSIS

TABLE 2: HEAVY METAL ANALYSIS

TABLE 3: BIOMASS ANALYSIS

TABLE 4: Porphyrin (Heme) ANALYSIS

TABLE 5: AMINO ACID COMPOSITION

Example IB: Identification of heme-overproducing algae

[0117] One of the identified strains was grown under fed-batch aerobic fermentation conditions where acetate is used as a reduced carbon source of nutrition for the culture. The strain was grown in a fermenter where minimal light can reach the culture. The strain was grown to a density that is greater than 120g/L and harvested via centrifugation. The harvested strain is red in color and can be added to compositions such as food products to confer a red, orange or brown color. FIG. 6 is a graph showing the ceil weight of the heme overproducer strain grown m aerobic fermentation conditions.

Example 1C: High density growth of heme-overproducing algae

[0118] Strains of Chlamydomonas that were previously selected for their ability to overexpress heme were grown to high density. To do this, a basal media containing media components that would allow the culture to achieve 120 grams per liter was developed. The strains are fresh water algae as such media components when solubilized with water were made not to exceed lOmS/cm. Cultures were then grown using an aerobic fed-batch fermentation process. Cultures were fed with a media containing acetate as a carbon source, ammonium hydroxide as a nitrogen source, and phosphoric acid as a phosphate source. Cultures were fed using a one sided acid pH-stat to maintain the pH at 6.8. As shown in FIG. 6, cultures were allowed to grow for 7 day and titers of l20g/L of biomass were achieved. Heme and protoporphyrin IX was quantified by using a heme quantification assay (Abnova KA1617). Heme and protoporphyrin were found to be greater than 5% of the biomass by weight. Titers of greater than lg/L of heme and protoporphyrin IX were achieved. In short, heme/protoporphyrin IX were extracted from a defined amount of algae culture by mixing the algae culture with a solution of 1.7M HCL and 80% Acetone. The mixture was allowed to sit for 30 minutes. After 30 minutes samples were centrifuge to separate the

heme/protoporphyrin IX extract from the algal biomass. The soluble heme/protoporphyrin IX samples were used in the assay from Abnova and compared to a standard curve to determine the amount of heme/protoporphyrin IX in the algal biomass. Example 2: Fractionation

[0119] Ceils from a heme overproducing strain of CMamydomonas reinhardtii were harvested from a fermentation culture. The harvested cells were disrupted by soni cation and then the samples were separated by centrifugation at 10,000 x G. This separated the samples into a carotenoid, starch and protein/heme biomass fractions. The protein/heme biomass was then re-suspended in Phosphate buffered saline pH 7.4. Shown in FIG. 4 is the fractionation following centrifugation (left) and the resuspension of the heme-containing fraction (right). Also shown in FIG. 5 illustrates process of PP1X and heme fractionation process and/or process of generating biomass, extracts, and/or lypophihzed products.

Example 3: Characterization of heme production

[0120] A number of heme assays can be used to determine the concentration of heme. In one example, the amount of heme can be quantitatively determined by mixing the algae biomass into an aqueous alkaline solution causing the heme to be converted into a uniform color. The intensity of the color can be measured by the absorbance at 400 nm which is directly proportional to the heme concentration in the sample. These measurements can then be compared to standards generated by heme at known concentrations to determine the amount of heme in algae samples.

Example 4: Preparation of a heme-enriched“meatless” burger

[0121] The heme-enriched samples can be used to prepare compositions of meat-like products produced from plant based materials and algae rich in heme. To create a heme- enriched burger, ingredients were mixed in the following proportions and formed into a disc shaped algae-plant based burger: 20% or about 20% Textured wheat protein, 20% or about 20% Refined coconut oil, 3% or about 3% Sunflower oil, 2% or about 2% Potato starch,

0.5% or about 0.5% Kojac gum, 0.5% or about 0.5% Xanthan gum, 45% or about 20%water and 4-9% or about 4-9% Flavors, including yeast extract, garlic powder, onion powder, salt, and heme-enriched (“red”) algae. Shown in FIG. 10 are burgers created with O.Olg, 0.1 g, l.Og, and 5.0g of the heme enriched algae.

[0122] In this example, the composition of the heme-enriched algae was 4.5%

protoporphyrin IX, 0.5% heme, 0% chlorophyll, 24.4% protein, 9% dietary fiber, 40% starch, 0.8% omega-3-fatty acids, 3.9% other fats, 7.5% moisture, and 8.4% ash. Example 5: Preparation of a heme-enriched plant-based burger

[0123] The heme-enriched samples can be used to prepare burger compositions from plant based materials and algae rich in heme. To create a heme-enriched plant-based burger, ingredients were mixed in the following proportions and formed into a disc: 20% or about 20% Textured soy protein, 20% or about 20% Refined coconut oil, 3% or about 3%

Sunflower oil, 2% or about 2% Potato starch, 1% or about 1% methylcellulose, 45% or about 45% water and 4-9% or about 4-9% Flavors, including yeast extract, garlic powder, onion powder, salt, and heme-enriched (“red”) algae. Shown in FIG. 11 are the ingredient mixes of the plant-based burger ingredients with no heme-enriched algae (far left), with the addition of heme-enriched algae (second from left), the ingredients with the addition of heme-enriched algae shaped into a burger before and after cooking (thirds from left and far right photos, respectively). As shown, the addition of the heme-enriched algae confers a red/red-like color (resembling a burger with animal blood) to the ingredient mix and to the burger, and this color undergoes a transition when cooked.

[0124] In this example, the composition of the heme-enriched algae was 4.5%

protoporphyrin IX, 0.5% heme, 0% chlorophyll, 24.4% protein, 9% dietary fiber, 40% starch, 0.8% omega-3-fatty acids, 3.9% other fats, 7.5% moisture, and 8.4% ash.

Example 6: Preparation of a heme-enriched meatless“tuna”

[0125] The heme-enriched samples can be used to prepare fish-like compositions, as shown in FIG. 12. To create a heme-enriched meatless“fish”, ingredients were mixed in the following proportions: 20% or about 20% Textured soy protein, 65% or about 65% water and 10% or about 10% Flavors and 5% or about 5% heme-enriched (“red”) algae. Shown in FIG. 12 is a square portion of the meatless“tuna.”

[0126] In this example, the composition of the heme-enriched algae was 4.5%

protoporphyrin IX, 0.5% heme, 0% chlorophyll, 24.4% protein, 9% dietary fiber, 40% starch, 0.8% omega-3-fatty acids, 3.9% other fats, 7.5% moisture, and 8.4% ash.

Example 7: Growth of heme-enriched algae strain on glucose

[0127] A heme-enriched algae strain was grown in a media with glucose as the sole carbon source. Briefly, as shown in FIG. 2, media was prepared in water, providing per liter of total volume 25g anhydrous glucose, 5g KNO3, 0.5275g KH2PO4, 0.3925g MgS04*7H20, 0.03l275g FeS04*7 H2O, 0.007l25g H3BO3, 0.002 CuS04, 0.002775g ZnS04, 0.002425g C0SO4, 0.00325g MnCl ₂ *4H ₂ 0, 0.001 l5g (NH ₄ )6Mq7q24*4H ₂ 0, and 0.0l735g CaCl. The media was adjusted to pH 7.0, autoclaved and had a final pH between 5.5 to 6.5 The algae strain was inoculated at a density of about 0. lg/L.

[0128] The culture was placed in a dark incubator (devoid of light) and grown at 30°C on a rotating shaker platform. Culture density (measured by dry cell weight) and residual glucose concentration in the media were measured daily. FIG. 7 shows the increase in dry cell weight over time and a concomitant decrease in residual glucose in the media. Dry cell weight in this experiment reached over 25g/L dry cell weight.

Example 8: Extraction of heme fraction from whole biomass

[0129] Using the heme-enriched algae (grown similarly to Example 1), a heme-enriched fraction was prepared. Approximately lOOg of algae biomass was mixed with a 1.0L of a solution containing 80% acetone and 20% 1.7M HCL for 30 minutes. The biomass was allowed to settle and then the aqueous layer was extracted (containing heme and

protoporphyrin IX) away from the solid into new container. Centrifugation was applied to the extracted aqueous layer or in some experiments, the sample was filtered with a filter having a molecular cutoff of 0.4lim. The res ulting aqueous fraction was neutralized with 10M NaOH. Then water was added at 100 ml per 100 ml of sample. Following this mixture, the heme and protoporphyrin IX became insoluble and fell out of solution. The solution was then centrifuged to collect the solids (containing the heme and protoporphyrin IX) and dried to form a red powxler. FIG. 5 shows the red-like colored fractions (containing the heme and protoporphyrin IX) collected through the steps of the procedure. From 160g of red algae biomass, 7.7g of PPIXheme was extracted.

Example 9: Removal of fatty acids from algae biomass to enrich for heme

[0130] Dry Chlamydomonas cells were mixed together with water ethanol and hexane in a ratio of 6:77: 17 . Samples were allowed to separate for 4 hours . The aqueous layer containing the fatty acids was then removed. The sample was then centrifuged to full separate the solid biomass layer from any remaining fatty acids. The biomass was then dried prior to further analysis. FIG. 8 shows a biochemical analysis of the algae biomass before and after the fatty acid extraction, demonstrating a greater than 10-fold reduction in fatty acid content after the extraction procedure.

Example 10: Targeted Modification of chlorophyll pathway to create heme-enriched strains

[0131] Guide RNAs (sgRNAs) can be designed against any of the sub-units of the magnesium chelatase gene to cause a deletion or an insertion that renders the protein complex non-functional. Once designed sgRNAs can be combined with the Cas9 protein by incubating them at 37°C to form ribonuclear proteins (RNPs). These RNPs carrying the sgRNAs to target magnesium chelatase are then electroporated into green algae cultures. 3xl0 ⁸ cells are placed into MAX efficiency transformation buffer reagent for algae (Thermo fisher scientific) and placed into a cuvette with a 0.2cm gap. The electroporation voltage is set to 250V and the pulse interval is set to l5ms. Once electroporated cells are recovered in growth media with 40mM sucrose added to improve recovery efficiency. Cells are then plated on growth media containing agar and grown in the dark due to the photosensitivity of the magnesium chelatase mutants. Once recovered the population can be pulled and struck out for individual colonies. Plates are again placed in the dark for 2 to 3 weeks. Mutants of Mg-chelatase can be identified by eye as they are not green. Mutants are then sequenced to ensure that target mutation was introduced.

Example 11: Modification of chlorophyll pathway to create heme-enriched strains that are improved for different meat imitations.

[0132] Strains of algae that increase the precursors to heme such as aminolevulinic acid can be mated to strains that are overexpressing heme to further increase the amount of heme or protoporphyrin IX that are produced. Mating can be done by identifying strains of Chlamydomonas that are the opposite mating type and then starving them for nitrogen. After nitrogen starvation, strains are re-suspended in water to promote the formation of flagella.

The flagella of the different mating types assist in the fusion of algae strains that will result in the formation of a zygote. The mated cultures are then exposed to chloroform to kill strains that did not mate. The chloroform does not kill zygotes. The zygotes are then placed into growth medium and allowed to propagate. Individual colonies are then identified and screened for an increase in heme by measuring for an increase in fluorescence of the precursor protoporphyrin IX or by biochemical assay (Abnova KA1617). [0133] Strains of algae overexpressing heme can also by mated with strains that are under or overproducing omega-3s, omega-6s or omega-9s. For imitation fish, more omega oils in strains of algae overexpressing heme are ideal. For imitation beef-like products, less omega oils in strains of algae overexpressing heme are ideal. As such strains of algae that are mutants for either over or underexpressing omega oils can be mated with strains of algae overexpressing heme to form a more ideal algae for various meat-like products.

SEQUENCES

ALA dehydratase (ALAD) nucleic acid sequence (SEQ ID NO: 1):

atgcagatgatgcagcgcaacgttgtgggccagcgccccgtcgctggctcccgccgc tcgctggtggttgccaac gttgcggaggtgacccgccccgcggtcagcaccaacggcaagcaccggactggtgtgccg gagggaactcccatc gtcacccctcaggacctgccctcgcgccctcgccgcaaccgccgcagcgagagcttccgt gcttccgttcgtgag gtgaacgtgtcgcccgccaacttcatcctgccgatcttcatccacgaggagagcaaccag aacgtgcccatcgcc tccatgcctggcatcaaccgcctggcgtatggcaagaacgtgattgactacgttgctgag gctcgctcttacggt gtcaaccaggtcgtggttttccccaagacgcccgaccacctgaagacgcaaaccgcggag gaggcgttcaacaag aacggcctcagccagcgcacgatccgcctgctgaaggactctttccctgacctggaggtg tacacggacgtggct ctggacccctacaactcggacggccacgacggtatcgtgtcggacgccggtgtgatcctg aacgacgagaccatc gagtacctgtgccgccaggccgtgagccaggccgaggccggtgccgacgtggtgtcgccc tctgacatgatggac ggccgcgtgggcgccatccgccgcgccctggaccgcgagggcttcaccaacgtgtccatc atgtcctacaccgcc aagtacgcctccgcctactacggccccttccgtgacgccctggcgtccgcgcccaagccc ggccaggcgcaccgc cgcatcccccccaacaagaagacctaccagatggaccccgccaactaccgcgaggccatc cgcgaggccaaggcc gacgaggccgagggcgctgacatcatgatggtcaagcccggcatgccgtacctggacgtg gtacgcctgctgcgt gagaccagcccgctgcccgtggccgtgtaccacgtgtcgggcgagtacgccatgctcaag gcggcggcggagcgc ggctggctgaacgagaaggatgccgtgcttgaggccatgacctgcttccgccgcgccggc gctgacctcatcctc acctactacggcattgaggcctccaagtggctggcgggcgagaagtaa

ALA dehydratase (ALAD) amino acid sequence (SEQ ID NO: 2):

MQMMQRNWGQRPVAGSRRSLWANVAEVTRPAVSTNGKHRTGVPEGTPIVTPQDLPSRPRR NRRSESFRASVRE VNVSPANFILPIFIHEESNQNVPIASMPGINRLAYGKNVIDYVAEARSYGVNQVWFPKTP DHLKTQTAEEAFNK NGLSQRTIRLLKDSFPDLEVYTDVALDPYNSDGHDGIVSDAGVILNDETIEYLCRQAVSQ AEAGADWSPSDMMD GRVGAIRRALDREGFTNVSIMSYTAKYASAYYGPFRDALASAPKPGQAHRRIPPNKKTYQ MDPANYREAIREAKA DEAEGADIMMVKPGMPYLDWRLLRETSPLPVAVYHVSGEYAMLKAAAERGWLNEKDAVLE AMTCFRRAGADLIL TYYGIEASKWLAGEK

coproporphyrinogen III oxidase (CPX1) nucleic acid sequence (SEQ ID NO: 3):

atggcactgcaagcctcaacccgctcgctccagcagcgccgcgccttctcttcggcc cagacctccaagcgtgtg tctgtgaccaaggtccgcgcgacggctatcgaggcggagaactatgtgaagcaggctccc cagtcgctggtccgc ccgggcatcgacactgaggactctatgcgcgctcgcttcgagaaggtgatccgcaacgcc caggactccatctgc aatgctatctccgagatcgatggcaagccgttccaccaggacgcctggacccgccccggc ggcggtggcggcatc agccgcgtgctgcaggacggcaacgtgtgggagaaggccggcgtcaacgtgtccgtggtc tacggcaccatgccc cctgaggcctaccgcgctgccactggcaacgccgagaagctgaagaacaagggtgacggt ggccgcgtgcccttc ttcgccgccggcatctcgtcggtgatgcacccccgcaacccccactgccccaccatgcac ttcaactaccgctac ttcgagactgaggagtggaacggcatccccggccagtggtggttcggcggcggcaccgac atcacccccagctat gtggtgcccgaggacatgaagcacttccacggcacctacaaggcggtgtgcgaccgccac gatcccgcttactac gagaagttccgcacctggtgcgatgagtacttcctcatcaagcaccgcggcgagcgccgc ggcctgggcggcatc ttcttcgatgacctgaacgaccgcaaccccgaggacatcctgaagttctcgaccgacgcc gtgaacaacgtggtg gaggcatactgccccatcatcaagaagcacatgaacgacccctacacccccgaggagaag gagtggcagcagatc cgccgcggccgctacgtggagttcaacctggtctatgaccgcggcaccaccttcggcctg aagaccggcggccgc attgagtcgatcctcatgtccatgccccagaccgcctcatggctgtacgaccaccagccc aaggccggctcgccc gaggccgagctgctcgacgcctgccgcaacccccgcgtctgggtgtaa coproporphyrinogen III oxidase (CPX1) amino acid sequence (SEQ ID NO: 4):

MALQASTRSLQQRRAFSSAQTSKRVSVTKVRATAIEAENYVKQAPQSLVRPGIDTEDSMR ARFEKVIRNAQDSIC

NAISEIDGKPFHQDAWTRPGGGGGISRVLQDGNVWEKAGVNVSWYGTMPPEAYRAAT GNAEKLKNKGDGGRVPF

FAAGISSVMHPRNPHCPTMHFNYRYFETEEWNGIPGQWWFGGGTDITPSYWPEDMKH FHGTYKAVCDRHDPAYY

EKFRTWCDEYFLIKHRGERRGLGGIFFDDLNDRNPEDILKFSTDAVNNWEAYCPIIK KHMNDPYTPEEKEWQQI

RRGRYVEFNLVYDRGTTFGLKTGGRIESILMSMPQTASWLYDHQPKAGSPEAELLDA CRNPRVWV

coproporphyrinogen III oxidase (CPX2) nucleic add sequence (SEQ ID NO: 5):

atgctgaggaagcagattggtggatctggccagcagcgggcgggcctccgacgggtg aaccaaggacctgcgcgt cggcggttggcaccctgccgcgtggcggcccccgtgcaaacctcgtcctccgtcgccaca ttcaatggcttcgtg gactacattcacggactccagaagaacattctgagcactgctgaggatctggagaacggc gagcggaagtttgtt gttgaccgctgggagcgcgacgccagcaaccccaacgccgggtatggcattacgtgcgtg cttgaggacgggaag gtgctggagaaggccgcagccaatatctcagtggtgcgcgggacgctgtcggcgcagcgc gcagtggccatgagc tcccgcggccgcagcagcatcgaccccaagggcgggcagccctacgccgcggccgccatg agcctagtgttccac agcgcgcacccgctcatccccacgctgcgcgcgacgtgcggttgttccaggtgggcgatg aggcgtggtacggcg gtggctgtgacctgacgcccaactacctagacgtggaggactcgcagtccttccaccgct actggaaggacgtgt gcggcaagtacaagccgggcctgtacaccgagctcaaggagtggtgcgacaggtacttct acatcccggcccgca aagagcaccgtggcattggcggcctgttctttgatgacatggccactgcggaggcgggct gcgatgtggaggcgt ttgtgcgggaagtgggagatggcatcctgccctgctggctgcccatcgtggcgcggcacc gtggccagcccttca cggagcagcagcggcaatggcagctgctgcgccgcggtcgctacatcgagttcaacctgc tgtacgaccgcggca tcaagttcggtctggacggcggccgcatcgagagcatcatggtgtcggcgccgccgctga tcgcgtggaagtaca acgtggtgccacagccgggcagccccgaggaggagatgctgaaggtgcttcagcagcccc gcgagtgggcctga coproporphyrinogen ill oxidase (CPX2) amino add sequence (SEQ ID NO: 6):

MLRKQIGGSGQQRAGLRRVNQGPARRRLAPCRVAAPVQTSSSVATFNGFVDYIHGLQKNI LSTAEDLENGERKFV

VDRWERDASNPNAGYGITCVLEDGKVLEKAAANISWRGTLSAQRAVAMSSRGRSSID PKGGQPYAAAAMSLVFH

SAHPLIPTLRADVRLFQVGDEAWYGGGCDLTPNYLDVEDSQSFHRYWKDVCGKYKPG LYTELKEWCDRYFYIPAR

KEHRGIGGLFFDDMATAEAGCDVEAFVREVGDGILPCWLPIVARHRGQPFTEQQRQW QLLRRGRYIEFNLLYDRG

IKFGLDGGRIESIMVSAPPLIAWKYNWPQPGSPEEEMLKVLQQPREWA

Ferrochelatase from Chlamydomonas reinhardtii nucleic acid sequence (SEQ ID NO: 7): atggcgtcgtttggattgatgcaaaggacggtgcactgtccccagcttgtggaggagcgg tgttcgccggtcgct ggctgctctggtcgtggcctgccagttatccagcggcaacggcgtggcgtgtgcagtgcc accaacggtgtccag cgagggcgtgtgctgcgccggacggccgcttcgaccgacgtggtctccttcgtggacccc aatgacattagaaaa cccgcagcagcagcagctggccctgcggtggataaggtcggcgttctgctgttaaacctt ggcgggcccgaaaag ctcgacgacgtcaagcctttcctgtataacctattcgccgacccagaaattattcgcctg ccagcggcagctcag ttcctgcagccgctgctcgcgacgatcatctccacgcttcgcgccccgaagagcgcggag ggctatgaggccatt ggcggtggtagcccgttgcgtaggattacagacgagcaggcggaggcgctggcggagtct ctgcgcgccaagggc caacctgcgaacgtgtacgtgggcatgcgctattggcacccctacacggaggaggcgctg gagcacattaaggcc gacggcgtcacgcgcctggtcatcctcccgctgtaccctcagttctccatctctaccagc ggctccagccttcga ctgcttgagtcgctcttcaagagcgacatcgcgctcaagtcgctgcggcacacggtcatc ccgtcctggtaccag cggcggggctacgtgagcgcgatggcggacctgattgtagaggagctgaagaagttccgg gacgtgcccagcgtg gagctgtttttctccgcgcacggcgtgcccaagtcctacgtggaggaggcgggcgaccca tacaaggaggagatg gaggagtgcgtgcggctcattacggacgaggtcaagcggcgcggcttcgccaacacgcac acgctggcctaccag agccgcgtgggccccgcggaatggctcaagccgtacacggatgagtccatcaaggagctg ggcaagcgcggcgtc aagtcgctgctggcggtgcccatcagctttgtcagcgagcacattgagacgttggaggag atcgacatggagtac cgcgagctggcggaggagagcggcatccgcaactggggccgcgtgccggcgctgaacacc aacgccgccttcatc gacgacctggcggacgcggtgatggaggcgctgccctacgtgggctgcctggccgggccg acagactcgctggtg ccgctgggcgacctggagatgctgctgcaggcctacgaccgcgagcgccgcacgctgccg tcaccggtggtgatg tgggagtggggctggaccaagagcgcggagacgtggaacggccgcattgccatgattgcc atcatcatcatcctg gcgctggaggcagccagcggccagtccatcctcaaaaacctgttcctggcggagtag

Ferrochelatase from Chlamydomonas reinhardtii amino acid sequence (SEQ ID NO: 8) MASFGLMQRTVHCPQLVEERCSPVAGCSGRGLPVIQRQRRGVCSATNGVQRGRVLRRTAA STDWSFVDPNDIRK PAAAAAGPAVDKVGVLLLNLGGPEKLDDVKPFLYNLFADPEIIRLPAAAQFLQPLLATII STLRAPKSAEGYEAI GGGSPLRRITDEQAEALAESLRAKGQPANVYVGMRYWHPYTEEALEHIKADGVTRLVILP LYPQFSISTSGSSLR LLESLFKSDIALKSLRHTVIPSWYQRRGYVSAMADLIVEELKKFRDVPSVELFFSAHGVP KSYVEEAGDPYKEEM EECVRLITDEVKRRGFANTHTLAYQSRVGPAEWLKPYTDESIKELGKRGVKSLLAVPISF VSEHIETLEEIDMEY RELAEESGIRNWGRVPALNTNAAFIDDLADAVMEALPYVGCLAGPTDSLVPLGDLEMLLQ AYDRERRTLPSPWW EWGWTKSAETWNGRIAMIAI II ILALEAASGQSILKNLFLAE

Glutamate-l-semialdehyde aminotransferase (GSA) nucleic acid sequence (SEQ ID NO:

9):

atgcagatgcagctgaacgccaagaccgtgcagggcgccttcaaggcgcagcgccct cgctctgtccgcggcaac gtggcggtgcgcgcagtggccgctccccctaagctggtcaccaagcgctccgaggagatc ttcaaggaggctcag gagctgctgcccggtggcgtgaactcgcccgtgcgcgctttccgctcggttggtggcggc cccatcgtcttcgac agggtcaagggtgcctactgctgggacgtcgatggcaacaagtacatcgactacgttggc tcttggggccctgcc atttgcggccacggcaacgacgaggtcaacaacgccctgaaggcgcagatcgacaagggc acctcgttcggtgct ccctgcgagctggagaacgtgctggccaagatggtgattgaccgcgtgccctcggtggag atggtgcgcttcgtg tcctcgggcactgaggcgtgcctgtcggtgctgcgcctgatgcgcgcatacaccggccgc gagaaggtgctgaag ttcaccggctgctaccacggccacgccgactccttcctggtgaaggccggctccggtgtg atcaccctgggcctg cccgactcgcccggtgtgcccaagagcaccgccgccgccaccctgaccgccacctacaac aacctggactccgtg cgcgagctgttcgccgccaacaagggcgagattgccggtgtgatcctggagcccgtggtc ggcaacagcggcttc attgtgcccaccaaggagttcctgcagggcctgcgcgagatctgcacggctgagggcgcc gtgctgtgcttcgat gaggtcatgaccggcttccgcattgccaagggctgcgcccaggagcacttcggtatcacc cccgacctgaccacc atgggcaaggtcattggtggcggcatgcctgtgggcgcctacggcggcaagaaggagatc atgaagatggtcgcc cccgccggccccatgtaccaggccggcaccctttcgggcaaccccatggccatgactgcc ggcatcaagacgctg gagatcctgggccgccccggcgcctacgagcacctggagaaggtgaccaagcgcctgatc gacggcatcatggcc gccgccaaggagcacagccacgagatcaccggcggcaacatcagcggcatgtttggcttc ttcttctgcaagggc cctgtgacctgcttcgaggacgccctggcggccgacactgccaagttcgcgcgcttccac cgcggcatgctggag gagggcgtctacctggctccctcgcagttcgaggccggcttcacctctctggcccactcc gaggcggacgtggat gccacgatcgccgccgctcgccgcgtgttcgcccgcatctaa

Glutamate-l-semialdehyde aminotransferase (GSA) amino acid sequence (SEQ ID NO:

10):

MQMQLNAKTVQGAFKAQRPRSVRGNVAVRAVAAPPKLVTKRSEEIFKEAQELLPGGVNSP VRAFRSVGGGPIVFD RVKGAYCWDVDGNKYIDYVGSWGPAICGHGNDEVNNALKAQIDKGTSFGAPCELENVLAK MVIDRVPSVEMVRFV SSGTEACLSVLRLMRAYTGREKVLKFTGCYHGHADSFLVKAGSGVITLGLPDSPGVPKST AAATLTATYNNLDSV RELFAANKGEIAGVILEPWGNSGFIVPTKEFLQGLREICTAEGAVLCFDEVMTGFRIAKG CAQEHFGITPDLTT MGKVIGGGMPVGAYGGKKEIMKMVAPAGPMYQAGTLSGNPMAMTAGIKTLEILGRPGAYE HLEKVTKRLIDGIMA AAKEHSHEITGGNISGMFGFFFCKGPVTCFEDALAADTAKFARFHRGMLEEGVYLAPSQF EAGFTSLAHSEADVD ATIAAARRVFARI

glutamyl-tma reductase (HEMA) nucleic acid sequence (SEQ ID NO: 11):

atgcagaccactatgcagcagcgtctccagggccgtaacgtggccgggcggagcgtc gctccctcggtccctgcc catcgctccttccactcacaccgggctgccactcaaaccgctacgatcagcgctgctgct agctcaaccaccaag ctgccagcttcgcatctggagagcagcaagaaggcgctggattcgctgaagcagcaggcc gtcaatcgctacgcg ggtgacaagaagagctccattattgccattggtctcaccattcacaacgcacccgtggag ctgcgcgagaagctg gctgtgcctgaggctgaatggccgcgtgctattgaggagctctgccagttcccgcacatc gaggaggccgcggtg ctgtcgacgtgcaatcgcatggagctctacgttgtcggtctgtcgtggcaccgcggcgtt cgcgaggtggaggag tggctgtctcgcaccagcggcgtgcctctggatgagctgcgcccctacctgttcctgctg cgcgaccgcgacgcc acgcaccacctgatgcgcgtgtcgggtggccttgactcgctggttatgggcgagggccag attctcgcccaagtg cgccaggtctacaaggtcggccagaactgccccggcttcggtcgccacctgaacggcctg ttcaagcaggctatc accgctggcaagcgcgtgcgtgccgagacctccatctccaccggctccgtctccgtctca tccgccgccgtcgag ctggcgcagctcaagctccccacccacaactggtccgacgctaaggtctgcatcatcggc gctggcaagatgtct acgctgctggtgaagcacctgcagagcaagggctgcaaggaggtgacggtgctcaaccgc tctctgccgcgcgcc caggcgctggcggaggagttccctgaggtcaagttcaacatccacctgatgcccgacctg ctgcagtgcgtggag gccagcgacgtcatcttcgccgcctccggctctgaggagatcctcatccacaaggagcat gtcgaggccatgtcc aagccatcggacgttgttggctccaagcgccgcttcgtcgacatctccgtgccccgcaac atcgcccccgccatc aacgagctggagcacggcatcgtctacaacgtcgacgacctgaaggaggttgtggccgcc aacaaggagggccgc gcgcaggcggccgccgaggccgaggtgctgatccgcgaggagcagcgcgcgttcgaggcc tggcgtgactctctg gagaccgtgcccaccatcaaggcgctgcgctccaaggccgagaccatccgcgccgccgag tttgagaaggccgtg tctcgcctgggcgaggggctatccaagaagcagctcaaggcggtggaggagctcagcaag ggcatcgtcaacaag ctgctgcacgggcccatgacggcactgcgctgcgacggcaccgatccggatgccgtgggc cagaccctcgcgaac atggaggccctggagcgcatgttccagctctcggaggtggacgtggccgcgctggcgggc aagcagtaa glutamyl-tma reductase (HEMA) amino acid sequence (SEQ ID NO: 12):

MQTTMQQRLQGRNVAGRSVAPSVPAHRSFHSHRAATQTATISAAASSTTKLPASHLESSK KALDSLKQQAVNRYA GDKKSSIIAIGLTIHNAPVELREKLAVPEAEWPRAIEELCQFPHIEEAAVLSTCNRMELY WGLSWHRGVREVEE WLSRTSGVPLDELRPYLFLLRDRDATHHLMRVSGGLDSLVMGEGQILAQVRQVYKVGQNC PGFGRHLNGLFKQAI TAGKRVRAETSISTGSVSVSSAAVELAQLKLPTHNWSDAKVCI IGAGKMSTLLVKHLQSKGCKEVTVLNRSLPRA QALAEEFPEVKFNIHLMPDLLQCVEASDVIFAASGSEEILIHKEHVEAMSKPSDWGSKRR FVDISVPRNIAPAI NELEHGIVYNVDDLKEWAANKEGRAQAAAEAEVLIREEQRAFEAWRDSLETVPTIKALRS KAETIRAAEFEKAV SRLGEGLSKKQLKAVEELSKGIVNKLLHGPMTALRCDGTDPDAVGQTLANMEALERMFQL SEVDVAALAGKQ

Light independent protochlorophyllide reductase subunit N (chlN) nucleic acid sequence (SEQ ID NO: 13):

atgttatactcacaatttaaacattcggtgcctttaggccgtaagtctccccttctt tcagggggccccccttct gggggtcgcccaacaacggctgcctcaggcctaggtcgcaacgtggccgtaagaattggg accccgttgggcttt gcccttcgggcccaggtaattatggcagctgcgggcaatactagcggtgcgccgcacccc gtaggggagtcccag cctgcgttgtcccaggtggattctcaacttgtaattgagtgtgaaacaggaaattaccat actttttgcccaatt agttgtgtttcttggttataccaaaaaattgaagatagttttttcttagttattggtaca aaaacgtgtgggtat tttttacaaaatgctttaggggttatgatttttgccgaacctcgttacgctatggcggaa ttagaagaaagcgat atttcggcgcaattaaatgattacaaagaattaaaacgtctatgtttacaaattaaacaa gaccgtaacccaagt gttattgtgtggattggcacatgcacaaccgaaattattaaaatggatttagaaggtatg gcaccgaaactagaa gctgaaatcggtattccaattgtggtagcacgcgcaaatggacttgattatgcttttaca caaggtgaagatact gttttagctgcgatggtccaaaaatgcccggaattaggcgctattccagctattgtacct cagattccttctgac tctcgtacacttagccaactatctgtagcggcttcggtacccgaaaacagtgcgtctggg ccagaaggggagcct tcactagcccagaagggaatggattctaagttaacaaacaactctccatgccgagtagat tctgtctcagaatct accccggcgtttcctggacgtgctccgcacgtcgggaaaagtactcctcaaaatttagtt ttatttggttcatta cctagcacgatggcaaatcaactggagtttgaattaaaacgccaaggtattaatgttact gggtggttacctgcg gctcgctattcatctttacctgcattaggtgaaaacgtgtatgtttgtgggattaatcca tttttaagtcgaact gctacttctttaatgcgtcgtcgtaaatgcaaattaatttcagctcctttcccaattggt ccagatggtacaaaa gcttgggtcgaaaaaatttgtaatgttttcggtgttacaccaactggtttagaagatcgt gaacgtcttgtttgg gaaggtttaaaagattatttaaatttcgtaaaagggaaatctgttttctttatgggtgat aatctgttagaaatt tcattagcccgttttttaattcgctgtggtatgaccgtttatgaaatcggtattccgtac atggaccaacgattt caagctggggaattagaattattaaaaaaaacatgcatggaaatgaacgtgcccctaccg cgtattgttgaaaaa cctgataattactatcaaattcaacgtattaaagaattacaaccagatttagttattacc ggcatggcccatgca aacccactggaagcgcgcggcattactacgaaatggtccgttgaatttacgtttgcgcaa attcatgggtttggc aacgcacgtgatatcttagaattagttacaaaaccgttacgtcgtaataaaaatctatct aaatatcaatttccg ttagatagctgggacaagcctgcttccgtaggcgctcacgaactgtcggcctaa

Light independent protochlorophyllide reductase subunit N (chlN) amino acid sequence (SEQ ID NO: 14):

MLYSQFKHSVPLGRKSPLLSGGPPSGGRPTTAASGLGRNVAVRIGTPLGFALRAQVIMAA AGNTSGAPHPVGESQ PALSQVDSQLVIECETGNYHTFCPISCVSWLYQKIEDSFFLVIGTKTCGYFLQNALGVMI FAEPRYAMAELEESD ISAQLNDYKELKRLCLQIKQDRNPSVIVWIGTCTTEI IKMDLEGMAPKLEAEIGIPIVVARANGLDYAFTQGEDT VLAAMVQKCPELGAIPAIVPQIPSDSRTLSQLSVAASVPENSASGPEGEPSLAQKGMDSK LTNNSPCRVDSVSES TPAFPGRAPHVGKSTPQNLVLFGSLPSTMANQLEFELKRQGINVTGWLPAARYSSLPALG ENVYVCGINPFLSRT ATSLMRRRKCKLISAPFPIGPDGTKAWVEKICNVFGVTPTGLEDRERLVWEGLKDYLNFV KGKSVFFMGDNLLEI SLARFLIRCGMTVYEIGIPYMDQRFQAGELELLKKTCMEMNVPLPRIVEKPDNYYQIQRI KELQPDLVITGMAHA NPLEARGITTKWSVEFTFAQIHGFGNARDILELVTKPLRRNKNLSKYQFPLDSWDKPASV GAHELSA Light Independent protochlorophyllide subunit B (chlB) nucleic acid sequence (SEQ ID NO: 15):

atgaaattagcgtattggatgtatgcgggaccggctcatattggaacattacgagtt gcaagctcgtttcgaaat gtgcatgctattatgcatgctcccttaggcgatgattattttaacgtaatgcgttcaatg ttagaacgtgaacgt gattttacgccagtgacggcaagtattgttgatcgtcatgttttagctcgtggttcacaa gaaaaagttgttgaa aacattcaacgaaaagataaagaagaatgtccggatttaattttattaacaccaacatgt acctcaagtattttg caagaagatttacaaaattttgtaaatcgcgcggccgaagtagcaaagcgttcggatgtt ttattagctgacgtt aaccattaccgagtgaatgaattacaagcggctgaccgtacgttagagcaaattgtacgc ttttatttagaaaaa gaagtaaataaacttcacgcggagttaggcggccttaaaaaaccgcttcgctttgcccag cgtacccaaaagccg tctgccaatattttaggcatgtttacactaggtttccataatcaacatgactgtcgtgaa ttaaaacgtttatta aatgatttaggtatcgaagtcaatgaagtgattcctgaaggtagttttgtacatggatta aaaaatttaccaaaa gcgtggtttaacatcgtcccgtatcgtgaagttggtttaatgacggcaatttatttagaa aaagaatttggcatg ccttatacctcaatcacgccaatgggcattattgacaccgcggcgtttattcgtgaaatt gcggccatttgtagt caaattagcacttcacaggcatctacaaactcaactgaaggactccagaggggagaaaat gtcagtttaactgaa actaattcgattatttttaataaagcaaaatatgaacaatacattaatcaacaaacgcat tttgtttctcaagca gcttggttttcacgttctattgactgtcaaaatttaaccggtaaaaaaaccgttgtgttt ggtgatgcaactcac gcggcaagtatgacgaaaattcttgtgcgcgaaatgggtattcatgttgtttgcgcgggc acgtattgtaaacat gatgcagattggtttagagagcaagtttcaggtttttgtgatcaagttttaattacagat gatcacagccaaatt gcggaaatcattgctcaaattgaacctgcagccatttttggtacacaaatggaacgtcat gttgggaaaaggtta gatattccttgtggggttatttctgcaccggtacatattcaaaacttcccactaggcttt agaccgtttttaggg tatgaaggtactaatcaaatttccgatttagtttataattcgtttagtttaggtatggaa gatcacttactagaa attttcaacggtcatgacaataaagaagttattacacgttcgtattcttcagaaactgat ttagaatggacaaaa gaagcattagatgaactagctcgtgttcctggttttgttcgttcaaaagttaaacgtaat actgaaaaatttgcg cgtacaaataaaaatcaagttattactattgaagttatgtacgcagctaaagaagcggta tcagcgtaa

Light Independent protochlorophyllide subunit B (chlB) amino acid sequence (SEQ ID NO: 16):

MKLAYWMYAGPAHIGTLRVASSFRNVHAIMHAPLGDDYFNVMRSMLERERDFTPVTASIV DRHVLARGSQEKWE NIQRKDKEECPDLILLTPTCTSSILQEDLQNFVNRAAEVAKRSDVLLADVNHYRVNELQA ADRTLEQIVRFYLEK EVNKLHAELGGLKKPLRFAQRTQKPSANILGMFTLGFHNQHDCRELKRLLNDLGIEVNEV IPEGSFVHGLKNLPK AWFNIVPYREVGLMTAIYLEKEFGMPYTSITPMGI IDTAAFIREIAAICSQISTSQASTNSTEGLQRGENVSLTE TNSI IFNKAKYEQYINQQTHFVSQAAWFSRSIDCQNLTGKKTVVFGDATHAASMTKILVREMGI HWCAGTYCKH DADWFREQVSGFCDQVLITDDHSQIAEI IAQIEPAAIFGTQMERHVGKRLDIPCGVISAPVHIQNFPLGFRPFLG YEGTNQISDLVYNSFSLGMEDHLLEIFNGHDNKEVITRSYSSETDLEWTKEALDELARVP GFVRSKVKRNTEKFA RTNKNQVITIEVMYAAKEAVSA

Light independent protochlorophyllide reductase subunit L (chlL) nucleic acid sequence (SEQ ID NO: 17):

atgaaattagcagtttatggcaaaggtggtattggtaaatccacaacaagttgtaac atttcaattgcattagca aaacgtggcaaaaaagtattacaaattggttgtgatccaaaacacgatagtacttttaca ttaaccggtttttta attccaacaattattgatactttacaaagtaaagattatcattacgaagatgtttggccg gaagatgttatttac caaggctacgggagtgtggattgtgttgaagcaggtggcccgccagccggcgccggctgt ggtgggtatgttgtt ggtgaaacagttaaattattaaaagaattaaatgcattttatgaatatgatgttattctg tttgatgttttaggg gatgttgtatgtggtgggtttgctgcacctttaaattacgccgactattgcattattgtc acagataatggcttt gatgcgttatttgccgcaaaccgtattgctgcttcagtgcgcgaaaaagcgcgcattcac ccattacgtttagct gggttaattgggaatcgtacagccaaacgcgatttaatcgataaatacgttgaagcgtgc ccgatgccagtctta gaggtattaccgttaattgaagacattcgtgtgtcacgcgtaaaaggtaaaacattattt gaaatggcagaacat gattcatcattacactacatttgtgacttttatttaaatattgcggatcaattattaact gaaccagaaggtgtt gttccgcgcgaattagcagaccgtgaattatttactctattatcagatttctatttaaac gctgggactcctagc cctagtggatctgagttcggctcaggcgcccttagcggaacgagcggcgaaacagctccc ggtaatatgggtcag cacatgagtaacgcagtaaaaacaaacgaacaggaaatgaatttctttcttgtgtaa Light independent protochlorophyllide reductase subunit L (chlL) amino acid sequence (SEQ ID NO: 18):

MKLAVYGKGGIGKSTTSCNISIALAKRGKKVLQIGCDPKHDSTFTLTGFLIPTI IDTLQSKDYHYEDVWPEDVIY QGYGSVDCVEAGGPPAGAGCGGYWGETVKLLKELNAFYEYDVILFDVLGDWCGGFAAPLN YADYCI IVTDNGF DALFAANRIAASVREKARIHPLRLAGLIGNRTAKRDLIDKYVEACPMPVLEVLPLIEDIR VSRVKGKTLFEMAEH DSSLHYICDFYLNIADQLLTEPEGWPRELADRELFTLLSDFYLNAGTPSPSGSEFGSGAL SGTSGETAPGNMGQ HMSNAVKTNEQEMNFFLV

Magnesium Chelatase subunit H (CHLH2) nucleic acid sequence (SEQ ID NO: 19): atgcggattgtgctggtcagcggcttcgagagctttaacgtgggcctgtacaaggatgcg gcggagctgctgaag cgctccatgcccaacgtcacactccaggtgttctccgaccgcgacctggcctccgacgcc acccgctcccggctg gaggcggctctggggcgcgccgacatcttcttcggatcactgctgttcgactacgaccag gtggagtggctacgg gcccggctggagcgggtgcctgtgcggctagtgtttgagtcggcgttggagctcatgagc tgcaacaaggtgggg tcgttcatgatgggcggcggcggtcccggcggcggcccgcccggcaaggcgcccggcccg ccgcccgcggtgaag aaggttctctccatgtttggaagcggtcgcgaggaggacaagatgggcggctcctccaat gtggtggccatgttc agttacctggtggagaccctgatggagccaacgggtgggttatttggtagttggtggttg tgttatggttggccg tttcggttgggtgatctgggctggtatctacaacccccctcaaccctcacgcctccaggc tacgtgccgccgcct gtggtggagactcccgcactgggctgcctccacccctccgcgcccggccgctacttcgag tcccccgccgagtac atgaagtggtacgccagggagggcccgctgcgcggcacgggcgccccggtggttggcgtg ctgctgtaccgcaag catgtgatcaccgaccagccgtacatcccgcagctggtcagccagctggaggcggagggg ctgctgcccgtgccc atcttcatcaacggcgtggaggcgcacaccgtggttcgcgacctgctgacctccgtgcac gagcaggatctgctt gcacgcggcgagacgggcgccatcagccccaccctgaagcgggacgcggtcaaggtggac gcggtggtgagcacc attggcttcccgctggtgggcggccccgccggcaccatggagggcgggcggcaggcggag gtggccaaggccatc ctgggcgccaaggacgtgccgtacacggtggcggcgccgctgcttattcaggacatggag agctggagcagggac ggcgtggcgggtctccagagtgtggtgctgtactcgctgccggagctggacggcgcagtg gacacggtgccactg ggggggctggtgggggacgacatctacctggtgccggagcgggtgaagaagctggcgggg cggctcaagtcgtgg cgtacgacacgcactaagcatgcctctgtttgtgacgtccagcccctcccccccccgtct cccctctccaccctc cctctcccttcctctcccttcctctcactctccaccctcttccccctccgcccaaacata acgaggcgggggctg ctgggcgcaagcgggccctggagtacccgctgcgacctagctagtccaactccacccatc ccccaatgccgcaat agctttccggagatgagcacacacacacacacacacacacacacacacacacacacacac acacacacacacaca cgccacccacgcacacacacacacacacacgctccccccgctcgccacacccccatccca ccccacccgcaggag ctgctgacgtaccccgcggactggggcccggccgagtggggcccgctgccctacctgccc gaccccgacgtgctg gttcgccgcatggaggcgcagtggggcgagctgcgagcctaccgcggcctcaacacctcg gcgcgcggcatgttc caggagtacggggctgacgtggtcctgcacttcggcatgcacggcaccgtggagtggttg cctggggcgccgctg gggaacaacggcctcagctggagcgacgtgctgctcggcgagctgccaaacgtgtacgtg tacgctgccaacaac ccctccgagtccatcgtggcaaagcggcgcggctacggcaccatcgtcagccacaacgtg ccgccgtacgggcgg gcgggtctgtacaagcagctttccagcctcaaggagacgcttcaggagtaccgcgaggcc gcgcaggccgcacgt gcccgagcaggagccagcagcagcagcggcagtagcagcagtagcagtagcagcggcagt ggcagtagcagcagc agtgtggagctgcgggcggcgttggcaccggtgttcgacgcctacactgaccgcctgtat gcctacctgcagctg ctggaggggcggctgttcagcgaggggctacacgtactgggagcgccgccggcgccgccg caggtgggtggtttt cccgcgagcttccaacggtaccgtaaactgcccaactgcccaacttctccccaaacacag gaggctgtcaagatc cggaacctgctcatgcagaacacgcaggagctggacgggctgctcaagggcctgggtggg cgttacgtgcttccc gaggcgggcggcgacctgctgcgggacgggtcgggcgtgctgcccaccggccgcaacatc cacgcactggacccc taccgcatgccctcccccgccgccatggcccgtggggcggcggtggcggcggccattctt gagcagcaccgggcg gctaacagcggggcgtggcccgagacctgcgccgtcaacctgtgggggctggactccatc aagagcaagggcgag agtgtgggggtggtgctggcgctggtgggggcggtgccggtgcgcgagggtacgggccgc gtcgcgcgcttccaa ctggtgccgctgtcagagttgggccggccgcgtgtggacgtgctttgtaacatgagcggc atcttccgcgactcc ttccagaacgtggtggagctgctcgacgacctgtttgcaagggccgccgccgccgctgac gagccagatgacatg aacttcatcgccaaacacgcccgagccatggagaagcagggcctgtccgccacctcggcc cgcctgttctccaac ccggctggcgactacgggtcgatggtcaacgagcgagtggggcagggcagctgggccaac ggcgacgagctgggt gacacgtgggcggcccgcaacgccttcagctacggccgaggcaaggagcgaggcacggcg cggcccgaggtgctg caggcgctgctcaagaccacggaccggatcgtgcagcagatcgacagtgtggagtacggc ctgacagacatccag gagtactacgccaacacgggcgccctcaagagagccgccgaggtggccaaaggcgacccg ggccccggtggccgg cggccgcgcgtggggtgttccattgtggaggcctttggcggcgcgggcgcgggcgcgggc ggcgccggtggagcg ggcgtgccgccgcctcgcgagctggaggaggtgctgcgcctggagtaccgctcgaagctg ctcaaccccaagtgg gcccgggccatggcggcgcagggcagcggcggcgcctacgagatcagtcagcgcatgacg gcgttggtgggctgg ggcgccaccaccgatttcagggagggctgggtgtgggacccaggcgccatggacacgtat gtgggcgatgaggag atggccagcaagctcaagaagaacaacccgcaggcctttgccaacgtgctgcggcgcatg ctggaggcggcgggc cgcggcatgtggagccccaacaaggaccagctggcacagctcaagtcgctgtacagcgag atggacgaccagctg gagggggtgacg

Magnesium Chelatase subunit H (CHLH2) amino acid sequence (SEQ ID NO: 20):

MRIVLVSGFESFNVGLYKDAAELLKRSMPNVTLQVFSDRDLASDATRSRLEAALGRA DIFFGSLLFDYDQVEWLR

ARLERVPVRLVFESALELMSCNKVGSFMMGGGGPGGGPPGKAPGPPPAVKKVLSMFG SGREEDKMGGSSNWAMF

SYLVETLMEPTGGLFGSWWLCYGWPFRLGDLGWYLQPPSTLTPPGYVPPPWETPALG CLHPSAPGRYFESPAEY

MKWYAREGPLRGTGAPWGVLLYRKHVITDQPYIPQLVSQLEAEGLLPVPIFINGVEA HTWRDLLTSVHEQDLL

ARGETGAISPTLKRDAVKVDAWSTIGFPLVGGPAGTMEGGRQAEVAKAILGAKDVPY TVAAPLLIQDMESWSRD

GVAGLQSWLYSLPELDGAVDTVPLGGLVGDDIYLVPERVKKLAGRLKSWRTTRTKHA SVCDVQPLPPPSPLSTL

PLPSSPFLSLSTLFPLRPNITRRGLLGASGPWSTRCDLASPTPPIPQCRNSFPEMST HTHTHTHTHTHTHTHTHT

RHPRTHTHTHAPPARHTPIPPHPQELLTYPADWGPAEWGPLPYLPDPDVLVRRMEAQ WGELRAYRGLNTSARGMF

QEYGADWLHFGMHGTVEWLPGAPLGNNGLSWSDVLLGELPNVYVYAANNPSESIVAK RRGYGTIVSHNVPPYGR

AGLYKQLSSLKETLQEYREAAQAARARAGASSSSGSSSSSSSSGSGSSSSSVELRAA LAPVFDAYTDRLYAYLQL

LEGRLFSEGLHVLGAPPAPPQVGGFPASFQRYRKLPNCPTSPQTQEAVKIRNLLMQN TQELDGLLKGLGGRYVLP

EAGGDLLRDGSGVLPTGRNIHALDPYRMPSPAAMARGAAVAAAILEQHRAANSGAWP ETCAVNLWGLDSIKSKGE

SVGWLALVGAVPVREGTGRVARFQLVPLSELGRPRVDVLCNMSGIFRDSFQNWELLD DLFARAAAAADEPDDM

NFIAKHARAMEKQGLSATSARLFSNPAGDYGSMVNERVGQGSWANGDELGDTWAARN AFSYGRGKERGTARPEVL

QALLKTTDRIVQQIDSVEYGLTDIQEYYANTGALKRAAEVAKGDPGPGGRRPRVGCS IVEAFGGAGAGAGGAGGA

GVPPPRELEEVLRLEYRSKLLNPKWARAMAAQGSGGAYEISQRMTALVGWGATTDFR EGWWDPGAMDTYVGDEE

MASKLKKNNPQAFANVLRRMLEAAGRGMWSPNKDQLAQLKSLYSEMDDQLEGVT

Magnesium Chelatase subunit 1 (CHLI1) Chlamydomonas reinhardtii nucleic acid sequence (SEQ ID NO: 21):

atggccctgaacatgcgtgtttcctcttccaaggtcgctgccaagcagcagggccgc atctccgcggtgccggtt gtgtcgagcaaggtggcctcctccgcccgcgtggcccccttccagggcgctcccgtggcc gcgcagcgcgctgct ctgctggtgcgcgccgctgccgctactgaggtcaaggctgctgagggccgcactgagaag gagctgggccaggcc cgccccatcttccccttcaccgccatcgtgggccaggatgagatgaagctggcgctgatt ctgaacgtgatcgac cccaagatcggtggtgtcatgatcatgggcgaccgtggcactggcaagtccaccaccatt cgtgccctggcggat ctgctgcccgagatgcaggtggttgccaacgacccctttaactcggaccccaccgacccc gagctgatgagcgag gaggtgcgcaaccgcgtcaaggccggcgagcagctgcccgtgtcttccaagaagattccc atggtggacctgccc ctgggcgccactgaggaccgcgtgtgcggcaccatcgacatcgagaaggcgctgaccgag ggtgtcaaggcgttc gagcccggcctgctggccaaggccaaccgcggcatcctgtacgtggatgaggtcaacctg ctggacgaccacctg gtcgatgtgctgctggactcggccgcctccggctggaacaccgtggagcgcgagggtatc tccatcagccacccc gcccgcttcatcctggtcggctcgggcaaccccgaggagggtgagctgcgcccccagctg ctggatcgcttcggc atgcacgcccagatcggcaccgtcaaggacccccgcctgcgtgtgcagatcgtgtcgcag cgctcgaccttcgac gagaaccccgccgccttccgcaaggactacgaggccggccagatggcgctgacccagcgc atcgtggacgcgcgc aagctgctgaagcagggcgaggtcaactacgacttccgcgtcaagatcagccagatctgc tcggacctgaacgtg gacggcatccgcggcgacatcgtgaccaaccgcgccgccaaggccctggccgccttcgag ggccgcaccgaggtg acccccgaggacatctaccgtgtcattcccctgtgcctgcgccaccgcctccggaaagac cccctggctgagatc gacgacggtgaccgcgtgcgtgagatcttcaagcaggtgttcggcatggagtaa

Magnesium Chelatase subunit 1 (CHLI1) Chlamydomonas reinhardtii amino acid sequence (SEQ ID NO: 22):

MALNMRVSSSKVAAKQQGRISAVPWSSKVASSARVAPFQGAPVAAQRAALLVRAAAATEV KAAEGRTEKELGQA RPIFPFTAIVGQDEMKLALILNVIDPKIGGVMIMGDRGTGKSTTIRALADLLPEMQWAND PFNSDPTDPELMSE EVRNRVKAGEQLPVSSKKIPMVDLPLGATEDRVCGTIDIEKALTEGVKAFEPGLLAKANR GILYVDEVNLLDDHL VDVLLDSAASGWNTVEREGISISHPARFILVGSGNPEEGELRPQLLDRFGMHAQIGTVKD PRLRVQIVSQRSTFD ENPAAFRKDYEAGQMALTQRIVDARKLLKQGEVNYDFRVKISQICSDLNVDGIRGDIVTN RAAKALAAFEGRTEV TPEDIYRVIPLCLRHRLRKDPLAEIDDGDRVREIFKQVFGME

Magnesium Chelatase sunubitl (CHLI2) Chlamydomonas reinhardtii nucleic acid sequence (SEQ ID NO: 23): atgcagagtctccagggtcagcgcgcgttcactgcggtgcgccagggtcgggcgggtccc ctgcggactcgcctg gtcgtgcgctcgtctgttgccttgccatccacgaaagccgcgaagaagccgaacttcccg ttcgtcaagattcag ggccaggaggagatgaagcttgcactgctgctgaacgtggtcgaccccaacatcggcgga gtgcttattatgggt gaccgcggcactgccaagtcggtcgcggtccgcgccctggtggatatgcttcccgacatt gacgtggttgagggc gacgccttcaacagctcccccaccgaccccaagttcatgggccccgacaccctgcagcgc ttccgcaacggcgag aagctgcccaccgtccgcatgcggacccccctggtggagctgcctctgggcgccaccgag gaccgcatctgcggc accatcgacatcgagaaggcgctgacgcagggcatcaaggcctacgagcccggcctgctg gccaaggccaaccgc ggcatcctgtatgtggacgaggtgaacctgctggatgatggcctggttgatgtcgtgctg gactcgtcggctagc ggcctgaacactgtggagcgtgagggtgtgtccattgtgcaccctgcccgcttcatcatg attggctcaggcaac ccccaggagggtgagctgcgcccgcagctgctggatcgcttcggcatgagcgtcaacgtg gccacgctgcaggac accaagcagcgcacgcagctggtgctggaccggcttgcgtacgaggcggaccctgacgca tttgtggactcgtgc aaggccgagcagacggcgctcacggacaagctggaggcggcccgccagcgcctgcggtcc gtcaagatcagcgag gagctgcagatcctgatctcggacatttgctcgcgcctggatgtggatggcctgcgcggt gacattgtgatcaac cgcgccgccaaggcgcttgtggccttcgagggccgcaccgaggtgaccacgaatgacgtg gagcgcgtcatctcg ggctgcctcaaccaccgcctgcgcaaggacccgctggaccccattgacaacggcaccaag gtggccatcctgttc aagcgcatgaccgaccccgagatcatgaagcgcgaggaggaggccaagaagaagcgcgag gaggcggccgccaag gccaaggcggagggcaaggcggaccgccccacgggcgccaaggctggcgcctgggctggc ttgccccctcgtcgg taa

Magnesium Chelatase sunubitl (CHLI2) Chlamydomonas reinhardtii amino acid sequence (SEQ ID NO: 24):

MQSLQGQRAFTAVRQGRAGPLRTRLWRSSVALPSTKAAKKPNFPFVKIQGQEEMKLALLL NWDPNIGGVLIMG DRGTAKSVAVRALVDMLPDIDWEGDAFNSSPTDPKFMGPDTLQRFRNGEKLPTVRMRTPL VELPLGATEDRICG TIDIEKALTQGIKAYEPGLLAKANRGILYVDEVNLLDDGLVDVVLDSSASGLNTVEREGV SIVHPARFIMIGSGN PQEGELRPQLLDRFGMSVNVATLQDTKQRTQLVLDRLAYEADPDAFVDSCKAEQTALTDK LEAARQRLRSVKISE ELQILISDICSRLDVDGLRGDIVINRAAKALVAFEGRTEVTTNDVERVISGCLNHRLRKD PLDPIDNGTKVAILF KRMTDPEIMKREEEAKKKREEAAAKAKAEGKADRPTGAKAGAWAGLPPRR

Magnesium Chelatase subunit D (CHLD) Chlamydomonas reinhardtii nucleic acid sequence (SEQ ID NO: 25):

atgaagtctctctgccatgagctcgctggccccagcgttactgggtgcggccggcga agcctccggaaggctttc agcggtgccaagattgcgcaggtctctcgccccgctgtgcttaacagcgtgcagcgccaa cagcgtctcgcctgt tctgccgtggccgagctctccgctgctgagctgcgcgccatgaaggtgtctgaggaggac tccaagggcttcgat gcggatgtgtcgacccgcctggcccgctcgtaccctctggcggccgtggtgggccaggac aacatcaagcaggcg ctgctgctgggcgccgtggacaccgggctgggcggcatcgccatcgccggtcgccgcggt accgccaagtccatc atggctcgcggcctgcacgctctgctgccgcccattgaggtggtggagggcagcatctgc aacgccgaccccgag gacccccgctcctgggaggctggcctggctgagaagtatgcgggcggccctgtgaagacc aagatgcgctcggcg ccgtttgtgcagatccctctgggtgtgactgaggaccgcttggtgggcactgtggacatt gaggcgtccatgaag gagggcaagactgtgttccagcccggcctgctggctgaggcgcaccgcggcatcctgtac gtggacgagatcaac ctgctggatgacggcattgccaacctgctgctgtccatcctgtcggacggagtcaacgtg gtggagcgcgagggc atctccatcagccacccctgccggccgctgctgattgccacctacaaccccgaggagggc cctctgcgtgagcac ctgctggaccgcatcgccattggcctcagcgccgacgtccccagcaccagcgacgagcgc gtcaaggccattgac gcagccatccgcttccaggacaagccgcaggacactattgacgacaccgcggagctcacc gacgccctgcgcacc tcggtcatcctggctcgcgagtacctgaaggacgtgaccatcgcgccggagcaggtgacc tacattgtggaggag gcgcgccgcggcggagtccaggggcaccgcgcggagctgtacgcggtcaagtgtgccaag gcgtgtgcggctctg gagggccgtgagcgtgtgaacaaggatgacctgcgccaggccgtgcagctggtcatcctg ccgcgcgccaccatc ctggaccagcccccgcccgagcaggagcagcccccgccgccgcccccgccccctcccccg ccgccgccgcaggac caaatggaggacgaggaccaggaggagaaggaggacgagaaggaggaggaggagaaggag aacgaggaccaggac gagcccgagatccctcaggagttcatgtttgagtccgagggcgtcatcatggacccctcc atcctcatgttcgcg cagcagcagcagcgcgcgcagggccgctccggccgcgccaagacgctcatcttcagcgac gaccgcggccgctac atcaagcccatgctgcccaagggtgacaaggtcaagcgcctggcagtggacgccacgctt cgcgccgccgcgccc taccagaagattcgccggcagcaggccatcagcgagggcaaggtgcagcgcaaggtgtac gtggacaagccagac atgcgctccaagaagctggcccgcaaggccggtgcgctggtgatttttgttgtggacgcg tccggctccatggct ctgaaccgcatgagcgccgccaagggcgcctgcatgcgcctgctggctgagtcgtacacc agccgcgaccaggtg tgcctcatccccttctacggcgacaaggccgaggtgctgctgccgccctccaagtccatc gccatggcccgccgc cgcctggactcgctgccctgcggcggcggctcgccccttgcgcacggcctgtccacggcg gtacgtgtgggcatg caggccagccaggcgggcgaggtgggccgcgtcatgatggtgctcatcacggacggccgc gccaacgtcagcctg gccaagtccaacgaggaccccgaggcgctcaagcccgacgcgcccaagcccaccgccgac tcgctgaaggacgag gtgcgcgacatggccaagaaggccgcgtccgccggcatcaacgtgcttgtcattgacacg gagaacaagttcgtg agcaccggctttgcggaggagatctccaaggcagcgcagggcaagtactactacctgccc aacgccagcgacgcc gccatcgcggcggccgcgtccggcgccatggccgcggccaagggcggctactag

Magnesium Chelatase subunit D (CHLD) Chlamydomonas reinhardtii amino acid sequence (SEQ ID NO: 26):

MKSLCHELAGPSVTGCGRRSLRKAFSGAKIAQVSRPAVLNSVQRQQRLACSAVAELSAAE LRAMKVSEEDSKGFD ADVSTRLARSYPLAAWGQDNIKQALLLGAVDTGLGGIAIAGRRGTAKSIMARGLHALLPP IEWEGSICNADPE DPRSWEAGLAEKYAGGPVKTKMRSAPFVQIPLGVTEDRLVGTVDIEASMKEGKTVFQPGL LAEAHRGILYVDEIN LLDDGIANLLLSILSDGVNWEREGISISHPCRPLLIATYNPEEGPLREHLLDRIAIGLSA DVPSTSDERVKAID AAIRFQDKPQDTIDDTAELTDALRTSVILAREYLKDVTIAPEQVTYIVEEARRGGVQGHR AELYAVKCAKACAAL EGRERVNKDDLRQAVQLVILPRATILDQPPPEQEQPPPPPPPPPPPPPQDQMEDEDQEEK EDEKEEEEKENEDQD EPEIPQEFMFESEGVIMDPSILMFAQQQQRAQGRSGRAKTLIFSDDRGRYIKPMLPKGDK VKRLAVDATLRAAAP YQKIRRQQAISEGKVQRKVYVDKPDMRSKKLARKAGALVIFWDASGSMALNRMSAAKGAC MRLLAESYTSRDQV CLIPFYGDKAEVLLPPSKSIAMARRRLDSLPCGGGSPLAHGLSTAVRVGMQASQAGEVGR VMMVLITDGRANVSL AKSNEDPEALKPDAPKPTADSLKDEVRDMAKKAASAGINVLVIDTENKFVSTGFAEEISK AAQGKYYYLPNASDA AIAAAASGAMAAAKGGY

Magnesium Chelatase subunit H (CHLH1) Chlamydomonas reinhardtii nucleic acid sequence (SEQ ID NO: 27):

atgcagacttcctcgcttcttggccggcgcacggcccacccggctgcgggcgcgacg cccaagccggttgcgccc tcgccccgcgtggctagcacccgccaggtcgcgtgcaatgtggcgactggaccccggccg cccatgaccaccttc accggtggcaacaagggccctgctaagcagcaggtgtcgctggatctgcgcgacgagggc gctggcatgttcacc agcaccagcccggagatgcgccgtgtcgtccctgacgatgtgaagggtcgcgttaaggtg aaggttgtgtacgtg gtgctggaggcccagtaccagtcggccatcagcgctgcggtgaagaacatcaacgccaag aactccaaggtgtgc ttcgaggtggtgggctacctgctggaggagctgcgtgaccagaagaacctcgatatgctc aaggaggatgtggcc tctgccaacatcttcatcggctcgctcatcttcattgaggagcttgccgagaagattgtg gaggcggtgagcccc ctgcgcgagaagctggacgcgtgcctgatcttcccgtccatgccggcggtcatgaagctg aacaagctgggcacg ttttcgatggctcagctgggccagtcgaagtcggtgttctcggagttcatcaagtctgct cgcaagaacaacgac aacttcgaggagggcttgctgaagctggtgcgcaccctgcctaaggtgctgaagtatctg ccctcggacaaggcg caggacgccaagaacttcgtgaacagcctgcagtactggctgggcggtaactcggacaac ctggagaacctgctg ctgaacaccgtcagcaactacgtgcccgctctgaagggcgtggacttcagcgtggctgag cccaccgcctacccc gatgtgggtatctggcaccctctggcctcgggcatgtacgaggacctgaaggagtacctg aactggtacgacacc cgcaaggacatggtcttcgccaaggacgcccccgtcattggcctggtgctgcagcgctcg cacctggtgactggc gatgagggccactacagcggcgtggtcgctgagctggagagccgcggtgctaaggtcatc cccgtctttgccggt ggcctggacttctccgcccccgtcaagaagttcttctacgaccccctgggctctggccgc acgttcgtggacacc gttgtgtcgctgaccggcttcgcgctggtgggcggccccgcgcgccaggacgcgccgaag gccattgaggcgctg aagaacctgaacgtgccctacctggtgtcgctgccgctggtgttccagaccactgaggag tggctggacagcgag ctgggcgtgcaccccgtccaggtggctctgcaggttgccctgcccgagctggatggtgcc atggagcccatcgtg ttcgctggccgtgactcgaacaccggcaagtcgcactcgctgcccgaccgcatcgcttcg ctgtgcgctcgcgcc gtgaactgggccaacctgcgcaagaagcgcaacgccgagaagaagctggccgtcaccgtg ttcagcttcccccct gacaagggcaacgtcggcactgccgcctacctgaacgtgttcggctccatctaccgcgtg ctgaagaacctgcag cgcgagggctacgacgtgggcgccctgccgccctcggaggaggatctgatccagtcggtg ctgacccagaaggag gccaagttcaactcgaccgacctgcacatcgcctacaagatgaaggtggacgagtaccag aagctgtgcccttac gccgaggcgctggaggagaactggggcaagccccccggcaccctgaacaccaacggccag gagctgctggtgtac ggccgccagtacggcaacgtcttcatcggcgtgcagcccaccttcggctacgagggcgac ccgatgcgcctgctg ttctcgaagtcggccagcccccaccacggcttcgccgcctactacaccttcctggagaag atcttcaaggccgac gccgtgctgcacttcggcacccacggctcgctggagttcatgcccggcaagcaggtcggc atgtcgggtgtgtgc taccccgactcgctgatcggcaccatccccaacctctactactacgccgccaacaacccg tctgaggccaccatc gccaagcgccgctcgtacgccaacaccatttcgtacctgacgccgcctgccgagaacgcc ggcctgtacaagggc ctgaaggagctgaaggagctgatcagctcgtaccagggcatgcgtgagtctggccgcgcc gagcagatctgcgcc accatcattgagaccgccaagctgtgcaacctggaccgcgacgtgaccctgcccgacgct gacgccaaggacctg accatggacatgcgcgacagcgttgtgggccaggtgtaccgcaagctgatggagattgag tcccgcctgctgccc tgcggcctgcacgtggtgggctgcccgcccaccgccgaggaggccgtggccaccctggtc aacatcgctgagctg gaccgcccggacaacaacccccccatcaagggcatgcccggcatcctggcccgcgccatt ggtcgcgacatcgag tcgatttacagcggcaacaacaagggcgtcctggctgacgttgaccagctgcagcgcatc accgaggcctcccgc acctgcgtgcgcgagttcgtgaaggaccgcaccggcctgaacggccgcatcggcaccaac tggatcaccaacctg ctcaagttcaccggcttctacgtggacccctgggtgcgcggcctgcagaacggcgagttc gccagcgccaaccgc gaggagctgatcaccctgttcaactacctggagttctgcctgacccaggtggtcaaggac aacgagctgggcgcc ctggtagaggcgctgaacggccagtacgtcgagcccggccccggcggtgaccccatccgc aaccccaacgtgctg cccaccggcaagaacatccacgccctggaccctcagtcgattcccactcaggccgcgctg aagagcgcccgcctg gtggtggaccgcctgctggaccgcgagcgcgacaacaacggcggcaagtaccccgagacc atcgcgctggtgctg tggggcactgacaacatcaagacctacggcgagtcgctggcccaggtcatgatgatggtc ggtgtcaagcccgtg gccgacgccctgggccgcgtgaacaagctggaggtgatccctctggaggagctgggccgc ccccgcgtggacgtg gttgtcaactgctcgggtgtgttccgcgacctgttcgtgaaccagatgctgctgctggac cgcgccatcaagctg gcggccgagcaggacgagcccgatgagatgaacttcgtgcgcaagcacgccaagcagcag gcggcggagctgggc ctgcagagcctgcgcgacgcggccacccgtgtgttctccaacagctcgggctcctactcg tccaacgtcaacctg gcggtggagaacagcagctggagcgacgagtcgcagctgcaggagatgtacctgaagcgc aagtcgtacgccttc aactcggaccgccccggcgccggtggcgagatgcagcgcgacgtgttcgagacggccatg aagaccgtggacgtg accttccagaacctggactcgtccgagatctcgctgaccgatgtgtcgcactacttcgac tccgaccccaccaag ctggtggcgtcgctgcgcaacgacggccgcacccccaacgcctacatcgccgacaccacc accgccaacgcgcag gtccgcactctgggtgagaccgtgcgcctggacgcccgcaccaagctgctcaaccccaag tggtacgagggcatg cttgcctcgggctacgagggcgtgcgcgagatccagaagcgcatgaccaacaccatgggc tggtcggccacctcg ggcatggtggacaactgggtgtacgacgaggccaactcgaccttcatcgaggatgcggcc atggccgagcgcctg atgaacaccaaccccaacagcttccgcaagctggtggccaccttcctggaggccaacggc cgcggctactgggac gccaagcccgagcagctggagcgcctgcgccagctgtacatggacgtggaggacaagatt gagggcgtcgaataa

Magnesium Chelatase subunit H (CHLH1) Chlamydomonas reinhardtii amino acid sequence (SEQ ID NO: 28):

MQTSSLLGRRTAHPAAGATPKPVAPSPRVASTRQVACNVATGPRPPMTTFTGGNKGPAKQ QVSLDLRDEGAGMFT STSPEMRRWPDDVKGRVKVKVVYWLEAQYQSAISAAVKNINAKNSKVCFEWGYLLEELRD QKNLDMLKEDVA SANIFIGSLIFIEELAEKIVEAVSPLREKLDACLIFPSMPAVMKLNKLGTFSMAQLGQSK SVFSEFIKSARKNND NFEEGLLKLVRTLPKVLKYLPSDKAQDAKNFVNSLQYWLGGNSDNLENLLLNTVSNYVPA LKGVDFSVAEPTAYP DVGIWHPLASGMYEDLKEYLNWYDTRKDMVFAKDAPVIGLVLQRSHLVTGDEGHYSGWAE LESRGAKVIPVFAG GLDFSAPVKKFFYDPLGSGRTFVDTWSLTGFALVGGPARQDAPKAIEALKNLNVPYLVSL PLVFQTTEEWLDSE LGVHPVQVALQVALPELDGAMEPIVFAGRDSNTGKSHSLPDRIASLCARAVNWANLRKKR NAEKKLAVTVFSFPP DKGNVGTAAYLNVFGSIYRVLKNLQREGYDVGALPPSEEDLIQSVLTQKEAKFNSTDLHI AYKMKVDEYQKLCPY AEALEENWGKPPGTLNTNGQELLVYGRQYGNVFIGVQPTFGYEGDPMRLLFSKSASPHHG FAAYYTFLEKIFKAD AVLHFGTHGSLEFMPGKQVGMSGVCYPDSLIGTIPNLYYYAANNPSEATIAKRRSYANTI SYLTPPAENAGLYKG LKELKELISSYQGMRESGRAEQICATIIETAKLCNLDRDVTLPDADAKDLTMDMRDSWGQ VYRKLMEIESRLLP CGLHWGCPPTAEEAVATLVNIAELDRPDNNPPIKGMPGILARAIGRDIESIYSGNNKGVL ADVDQLQRITEASR TCVREFVKDRTGLNGRIGTNWITNLLKFTGFYVDPWVRGLQNGEFASANREELITLFNYL EFCLTQWKDNELGA LVEALNGQYVEPGPGGDPIRNPNVLPTGKNIHALDPQSIPTQAALKSARLWDRLLDRERD NNGGKYPETIALVL WGTDNIKTYGESLAQVMMMVGVKPVADALGRVNKLEVIPLEELGRPRVDVVVNCSGVFRD LFVNQMLLLDRAIKL AAEQDEPDEMNFVRKHAKQQAAELGLQSLRDAATRVFSNSSGSYSSNVNLAVENSSWSDE SQLQEMYLKRKSYAF NSDRPGAGGEMQRDVFETAMKTVDVTFQNLDSSEISLTDVSHYFDSDPTKLVASLRNDGR TPNAYIADTTTANAQ VRTLGETVRLDARTKLLNPKWYEGMLASGYEGVREIQKRMTNTMGWSATSGMVDNWVYDE ANSTFIEDAAMAERL MNTNPNSFRKLVATFLEANGRGYWDAKPEQLERLRQLYMDVEDKIEGVE

Photochlorophyllide reductase subunit B (chlB) nucleic acid sequence (SEQ ID NO:

29):

atgaaattagcttattggatgtacgcaggtcccgctcatatcggtgtgttgcgtgtt agcagctcttttaaaaat gtacatgccattatgcatgctcctttaggagatgattattttaatgtaatgcgttccatg ttagaacgtgaacgt gattttacaccagtaacagccagtattgtagatcgtcatgttttagcaagaggatcgcaa gaaaaagtggttgaa aatattacgcgaaaaaataaagaagaaactcctgatttaattttattaactcctacttgt acgtcaagcatttta caagaagatttacacaattttgttgaatcggcattagctaaaccagtacaaatagatgaa catgcagaccataaa gtaactcaacaaagtgcactttcaagtgtatcccctttactaccgcttgaagaaaataca ttaatagtaagtgaa ctagataagaagcttagcccgtctagcaagttgcatattaatatgcccaatatttgtatt cccgaaggagaaggg gaaggggagcagactaaaaattcaatttttgttaaatctgcaactttaacaaatttgtca gaagaggaactatta aatcaagaacatcataccaaaacaagaaatcactctgacgttattttagctgatgtaaac cattatcgtgtaaat gaattacaagctgcagatcgtactcttgaacaaattgtacgttattatatttctcaagca caaaaacaaaattgt ttaaacattactaaaacagccaaaccatctgtaaatattattggtatttttactttgggt tttcataatcaacat gattgtcgtgaattaaaacgtttatttaatgatttaggtattcaaatcaatgaaatcata cctgaaggcggaaat gtacacaacttaaaaaaattaccccaagcttggtttaattttgtgccctaccgtgaaatt ggcttaatgactgct atgtatttaaaatccgagtttaatatgccttacgtcgcaattactcctatgggattaatt gatacggctgcttgt attcgttcaatttgtaaaatcattacaactcaattattaaatcagacggctacagtgcag gagccatcaaaattt atttacccgaaggcgacgtcattagaacaaaccaatattctcgaaacctctcaaaaagaa actattcttaaagac aatccagatagcggaaataccctttctacaactgtagaagaaattgaaactttatttaat aaatatatcgatcaa caaactcgttttgtttcccaagcagcctggttttcacgttctattgactgtcaaaattta acaggtaaaaaagcc gtagttttcggagatgctacacattcagctgccatgacaaaattattagcacgtgaaatg ggtattaaggtttca tgcgctggaacttattgcaaacacgatgcggattggtttagagagcaagttagtgggttt tgtgatcaagtttta attaccgatgatcacacacaagtaggggatatgattgcacaattagaacctgcagccatt tttgggacacaaatg gaacgtcacgttggtaaacgtttagatattccatgtggtgttatatctgctcctgtgcat attcaaaactttccg ttaggttatcgaccttttttaggttatgaaggtacaaatcaaatagctgatttagtgtat aattcatttaatctt ggaatggaagaccatttattacaaatttttggaggtcatgattcagaaaacaattcgtca attgcaacgcatttg aatacaaataacgcaataaatttagcgccaggatatttacctgagggagaaggcagtagt agaacttcaaatgta gtgtctacaatttctagtgaaaaaaaagccattgtatggtctccagaaggtttagcagaa ttaaataaagtccca ggatttgttcgaggaaaagttaaacgtaatacggaaaaatatgctttacaaaaaaattgt tcgatgattactgta gaagttatgtatgcagcaaaagaagctttgtcggcttaa

Photochlorophyllide reductase subunit B (chlB) amino acid sequence (SEQ ID NO: 30):

MKLAYWMYAGPAHIGVLRVSSSFKNVHAIMHAPLGDDYFNVMRSMLERERDFTPVTA SIVDRHVLARGSQEKWE

NITRKNKEETPDLILLTPTCTSSILQEDLHNFVESALAKPVQIDEHADHKVTQQSAL SSVSPLLPLEENTLIVSE

LDKKLSPSSKLHINMPNICIPEGEGEGEQTKNSIFVKSATLTNLSEEELLNQEHHTK TRNHSDVILADVNHYRVN

ELQAADRTLEQIVRYYISQAQKQNCLNITKTAKPSVNIIGIFTLGFHNQHDCRELKR LFNDLGIQINEIIPEGGN

VHNLKKLPQAWFNFVPYREIGLMTAMYLKSEFNMPYVAITPMGLIDTAACIRSICKI ITTQLLNQTATVQEPSKF

IYPKATSLEQTNILETSQKETILKDNPDSGNTLSTTVEEIETLFNKYIDQQTRFVSQ AAWFSRSIDCQNLTGKKA

WFGDATHSAAMTKLLAREMGIKVSCAGTYCKHDADWFREQVSGFCDQVLITDDHTQV GDMIAQLEPAAIFGTQM

ERHVGKRLDIPCGVISAPVHIQNFPLGYRPFLGYEGTNQIADLVYNSFNLGMEDHLL QIFGGHDSENNSSIATHL

NTNNAINLAPGYLPEGEGSSRTSNWSTISSEKKAIVWSPEGLAELNKVPGFVRGKVK RNTEKYALQKNCSMITV

EVMYAAKEALSA

Photochlorophyllide reductase subunit L (chIL) nucleic acid sequence (SEQ ID NO:

31):

atgaaattagctgtttacggaaaaggtggtattggaaaatcaacgacaagttgtaat atttcgattgctttacga aaacgtggtaaaaaagtgttacaaattggttgtgatcctaaacatgatagtacttttaca ttgacagggttttta attccaaccattattgatacattaagttctaaagattatcattatgaagatatttggccc gaagatgttatttac ggaggttatgggggtgtagattgtgttgaagctggaggaccacctgccggtgcggggtgt ggtggttatgttgta ggtgaaacggtaaaacttttaaaagagttaaatgcttttttcgaatacgatgttatttta tttgatgttttaggt gatgttgtttgtggtggctttgctgctccattaaactacgctgattattgtattattgta actgataatggtttt gatgctttatttgctgcaaatcgtattgcagcttcagttcgtgaaaaagcacgtacacat ccattgcgtttagcg ggtttaatcggaaatcgtacatcaaaacgtgatttaattgataaatatgtagaagcttgt cctatgccagtatta gaagttttaccattaattgaagaaattcgtatttcacgtgttaaaggcaaaactttattt gaaatgtcaaataaa aataatatgacttcggctcatatggatggctctaaaggtgacaattctacagtaggagtg tcagaaactccatcg gaagattatatttgtaatttttatttaaatattgctgatcaattattaacagaaccagaa ggagttattccacgt gaattagcagataaagaactttttactcttttatcagatttctatcttaaaatttaa

Photochlorophyllide reductase subunit L (chIL) amino acid sequence (SEQ ID NO: 32):

MKLAVYGKGGIGKSTTSCNISIALRKRGKKVLQIGCDPKHDSTFTLTGFLIPTI IDTLSSKDYHYEDIWPEDVIY GGYGGVDCVEAGGPPAGAGCGGYWGETVKLLKELNAFFEYDVILFDVLGDWCGGFAAPLN YADYCI IVTDNGF DALFAANRIAASVREKARTHPLRLAGLIGNRTSKRDLIDKYVEACPMPVLEVLPLIEEIR ISRVKGKTLFEMSNK NNMTSAHMDGSKGDNSTVGVSETPSEDYICNFYLNIADQLLTEPEGVIPRELADKELFTL LSDFYLKI

Photochlorophyllide reductase subunit N (chlN) nucleic acid sequence (SEQ ID NO:

33):

atgttagatggtgccacaacgattttaaatttaaatagtttttttgaatgtgaaact ggcaattatcatactttt tgcccgattagctgtgtagcttggttatatcaaaaaatcgaagatagcttttttttagta attgggacaaaaaca tgtggttattttttacaaaatgcccttggagttatgatttttgccgaacctaggtatgct atggcagaattagaa gaaagtgatatttcagcacaattaaacgattataaagaattaaaacgtttatgtttacaa attaaacaagataga aatcccagcgttattgtttggattggaacttgtacaactgaaattatcaaaatggattta gaagggatggctcca cgtttagaaactgaaatcggcatacccattgttgtagcacgtgctaatggtttagattat gcttttacacaaggt gaagacacagttttatcagcaatggccttagcatccttaaaaaaagatgttcctttttta gtaggtaatactggg ttaacaaacaaccagcttctccttgaaaaatcaacttcttcagttaatgggacagacgga aaggaattacttaaa aaatctcttgtattatttggttccgtaccaagtacagttactacacaattaactttagaa ttaaaaaaagaaggt attaatgtatctggatggcttccatctgctaattataaagatttacctacttttaataaa gatacacttgtatgt ggtataaatccttttttaagtcgaacagctaccacgttaatgcgtcgtagtaagtgcaca ttaatttgtgcaccc tttccaataggccccgatggcacaagagtttggattgaaaaaatttgtggtgcttttggc attaatcctagtctt aatccaattactggtaatactaatttatatgatcgtgaacaaaaaattttcaacgggcta gaagattatttaaaa ttattacgtggaaaatctgtattttttatgggtgataatttattagaaatttctttagca cgttttttaacacgt tgtggtatgattgtttatgaaatcggaattccatatttagataaacgatttcaagcagca gaattagctttatta gaacaaacttgtaaagaaatgaatgtaccaatgccgcgcattgtagaaaaaccagataat tattatcaaattcga cgtatacgtgaattaaaacctgatttaacgattactggaatggcacatgcaaatccatta gaagctcgaggtatt acaacaaaatggtcagttgaatttacttttgctcaaattcatggatttactaatacacgt gaaattttagaatta gtaacacagcctcttagacgcaatctaatgtcaaatcaatctgtaaatgctatttcttaa

Photochlorophyllide reductase subunit N (chlN) amino acid sequence (SEQ ID NO: 34):

MLDGATTILNLNSFFECETGNYHTFCPISCVAWLYQKIEDSFFLVIGTKTCGYFLQN ALGVMIFAEPRYAMAELE ESDISAQLNDYKELKRLCLQIKQDRNPSVIVWIGTCTTEIIKMDLEGMAPRLETEIGIPI WARANGLDYAFTQG EDTVLSAMALASLKKDVPFLVGNTGLTNNQLLLEKSTSSVNGTDGKELLKKSLVLFGSVP STVTTQLTLELKKEG INVSGWLPSANYKDLPTFNKDTLVCGINPFLSRTATTLMRRSKCTLICAPFPIGPDGTRV WIEKICGAFGINPSL NPITGNTNLYDREQKIFNGLEDYLKLLRGKSVFFMGDNLLEISLARFLTRCGMIVYEIGI PYLDKRFQAAELALL EQTCKEMNVPMPRIVEKPDNYYQIRRIRELKPDLTITGMAHANPLEARGITTKWSVEFTF AQIHGFTNTREILEL VTQPLRRNLMSNQSVNAIS

Porphobilinogen deaminase (PBGD1) nucleic acid sequence (SEQ ID NO: 35):

atgcagcagtgcgttggccgctccgtccgcgctccgtccagcagggcggtcgcgccc aaggtcgctggcgctcgt gtcagccgccgcgtgtgccgcgtctatgcctccgctgttgctaccaagacggtgaagatt ggcacgcgcggctcg cccctggctctggcccaggcttacatgactcgcgacctgctgaagaagagcttccctgag ctgagcgaggagggt gctctggagatcgtgatcatcaagaccaccggtgacaaaatcctgaaccagcccctggct gacatcggtggcaag ggtctgtttaccaaggagatcgatgatgctctgctgagcggcaagattgacatcgccgtg cactccatgaaggac gtgcccacctacctgcccgagggcaccatcctgccctgcaacctgccccgcgaggatgtg cgcgatgtgttcatc tcgcctgtcgccaaggacctgagcgagctgcccgccggcgccattgtgggctcggcctcg ctgcgccgtcaggcc cagatcctggccaagtacccccacctcaaggtggagaacttccgcggcaacgtgcagacc cgcctgcgcaagctg aacgagggcgcctgctccgccaccctgctggctctggccggtctgaagcgcctggacatg actgagcacatcacc aagaccctcagcattgacgagatgctgcccgccgtgagccagggcgccattggcattgcc tgccgcaccgacgac ggcgccagccgcaacctgctggccgccctgaaccacgaggagacccgcatcgccgtggtg tgcgagcgcgccttc ctgaccgccctggacggctcttgccgcacccccattgccggctacgcgcacaagggcgcc gacggcatgctgcac ttcagcggcctggtggccaccccggacggcaagcagatcatgcgcgctagccgcgtggtg cccttcacggaggcg gatgccgtcaagtgcggcgaggaggccggcaaggagctcaaggccaacggccccaaggag ctgttcatgtactaa

Porphobilinogen deaminase (PBGD1) amino acid sequence (SEQ ID NO: 36):

MQQCVGRSVRAPSSRAVAPKVAGARVSRRVCRVYASAVATKTVKIGTRGSPLALAQAYMT RDLLKKSFPELSEEG ALEIVI IKTTGDKILNQPLADIGGKGLFTKEIDDALLSGKIDIAVHSMKDVPTYLPEGTILPCNLP REDVRDVFI SPVAKDLSELPAGAIVGSASLRRQAQILAKYPHLKVENFRGNVQTRLRKLNEGACSATLL ALAGLKRLDMTEHIT KTLSIDEMLPAVSQGAIGIACRTDDGASRNLLAALNHEETRIAWCERAFLTALDGSCRTP IAGYAHKGADGMLH FSGLVATPDGKQIMRASRWPFTEADAVKCGEEAGKELKANGPKELFMY

Porphobilinogen deaminase (PBGD2) nucleic acid sequence (SEQ ID NO: 37):

atgcgatcgtatctgctcaaggctcaagtggcctcatgtcagttttcgcgcacgtcg aaggtctggagactggcg ccgggttctgacagacgacggtgtcggggcctcactcggacaccgcactgcgcggccccc accagcgagcccgcc ccgccatccagcagcggcaagagcgggcaacgaccactcgtgatagccacgcggccatct aagcttgcaaaggag cagacgcggcaggtgcagcagctgctgctggcggcggcgcagctcaaggacgagcagctg cagctgagcaccctg gaactggcgtctaggggcgacacgactcagggtgtgtcgctgcgcagtctgggctcgggc gcattcaccgaggag ctggaccaggctgtgctgtcgggcgctgccgacatgtcggtgcacagcctgaaggactgc cccgccgccctggcg cccgggctgctgctggccgcctgcctgccgcgggccgacccccgggacgtcctcatcgcg cccgaggccacctcg ctgggcgagctggtgccgggcagccgtgtgggcaccagcagcagccgccgcgcggcgcag atcaagcactccttc ccccacctgcaggttgtgcagctgcgcggcaatgtggactcgcggctggggcgcatccgc agccgcgacatcggc gccacagtgctggcggcggcgggcctcaagcggctgggtgtgatgaactcggacgagggt gacactaccgctacg ggcgccgtgggggtggtgtgcagggcagacgatgagtgggtggtcggcctgctggacgcc atctcgcaccgcggc acggccctggaggtggcggcggagcgggcgtgcctggcagcgctgctgggcggcggcggc gcgtgccagcgttca gcgttcccggacattgcgtgggcctgccacacgcggcacgaccccgacagcaacacaatg gacctggattgcctg gtggcggacctggagggcaaggagctcttcaggtacacggagttctaccggccggtcatt gacgaggtggacgcg gtgtcgctggggtcgctgtacggcagcctgctgcgcatgatggcgccaccaggcgcggcc ccctgttggcagcta ccttcctcgcggcattag

Porphobilinogen deaminase (PBGD2) amino acid sequence (SEQ ID NO: 38):

MRSYLLKAQVASCQFSRTSKWRLAPGSDRRRCRGLTRTPHCAAPTSEPAPPSSSGKSGQR PLVIATRPSKLAKE

QTRQVQQLLLAAAQLKDEQLQLSTLELASRGDTTQGVSLRSLGSGAFTEELDQAVLS GAADMSVHSLKDCPAALA

PGLLLAACLPRADPRDVLIAPEATSLGELVPGSRVGTSSSRRAAQIKHSFPHLQWQL RGNVDSRLGRIRSRDIG

ATVLAAAGLKRLGVMNSDEGDTTATGAVGWCRADDEWWGLLDAISHRGTALEVAAER ACLAALLGGGGACQRS

AFPDIAWACHTRHDPDSNTMDLDCLVADLEGKELFRYTEFYRPVIDEVDAVSLGSLY GSLLRMMAPPGAAPCWQL

PSSRH

Protoporphyrinogen oxidase (PPX1) nucleic acid sequence (SEQ ID NO: 39):

atgatgttgacccagactcctgggaccgccacggcttctagccggcggtcgcagatc cgctcggctgcgcacgtc tccgccaaggtcgcgcctcggcccacgccattctcggtcgcgagccccgcgaccgctgcg agccccgcgaccgcg gcggcccgccgcacactccaccgcactgctgcggcggccactggtgctcccacggcgtcc ggagccggcgtcgcc aagacgctcgacaatgtgtatgacgtgatcgtggtcggtggaggtctctcgggcctggtg accggccaggccctg gcggctcagcacaaaattcagaacttccttgttacggaggctcgcgagcgcgtcggcggc aacattacgtccatg tcgggcgatggctacgtgtgggaggagggcccgaacagcttccagcccaacgatagcatg ctgcagattgcggtg gactctggctgcgagaaggaccttgtgttcggtgaccccacggctccccgcttcgtgtgg tgggagggcaagctg cgccccgtgccctcgggcctggacgccttcaccttcgacctcatgtccatccccggcaag atccgcgccgggctg ggcgccatcggcctcatcaacggagccatgccctccttcgaggagagtgtggagcagttc atccgccgcaacctg ggcgatgaggtgttcttccgcctgatcgagcccttctgctccggcgtgtacgcgggcgac ccctccaagctgtcc atgaaggcggccttcaacaggatctggattctggagaagaacggcggcagcctggtggga ggtgccatcaagctg ttccaggaacgccagtccaacccggccccgccgcgggacccgcgcctgccgcccaagccc aagggccagacggtg ggctcgttccgcaagggcctgaagatgctgccggacgccattgagcgcaacatccccgac aagatccgcgtgaac tggaagctggtgtctctgggccgcgaggcggacgggcggtacgggctggtgtacgacacg cccgagggccgtgtc aaggtgtttgcccgcgccgtggctctgaccgcgcccagctacgtggtggcggacctggtc aaggagcaggcgccc gccgccgccgaggccctgggctccttcgactacccgccggtgggcgccgtgacgctgtcg tacccgctgagcgcc gtgcgggaggagcgcaaggcctcggacgggtccgtgccgggcttcggtcagctgcacccg cgcacgcagggcatc accactctgggcaccatctacagctccagcctgttccccggccgcgcgcccgagggccac atgctgctgctcaac tacatcggcggcaccaccaaccgcggcatcgtcaaccagaccaccgagcagctggtggag caggtggacaaggac ctgcgcaacatggtcatcaagcccgacgcgcccaagccccgtgtggtgggcgtgcgcgtg tggccgcgcgccatc ccgcagttcaacctgggccacctggagcagctggacaaggcgcgcaaggcgctggacgcg gcggggctgcagggc gtgcacctggggggcaactacgtcagcggtgtggccctgggcaaggtggtggagcacggc tacgagtccgcagcc aacctggccaagagcgtgtccaaggccgcagtcaaggcctaa

Protoporphyrinogen oxidase (PPX1) amino acid sequence (SEQ ID NO: 40):

MMLTQTPGTATASSRRSQIRSAAHVSAKVAPRPTPFSVASPATAASPATAAARRTLHRTA AAATGAPTASGAGVA KTLDNVYDVIWGGGLSGLVTGQALAAQHKIQNFLVTEARERVGGNITSMSGDGYVWEEGP NSFQPNDSMLQIAV DSGCEKDLVFGDPTAPRFVWWEGKLRPVPSGLDAFTFDLMSIPGKIRAGLGAIGLINGAM PSFEESVEQFIRRNL GDEVFFRLIEPFCSGVYAGDPSKLSMKAAFNRIWILEKNGGSLVGGAIKLFQERQSNPAP PRDPRLPPKPKGQTV GSFRKGLKMLPDAIERNIPDKIRVNWKLVSLGREADGRYGLVYDTPEGRVKVFARAVALT APSYWADLVKEQAP AAAEALGSFDYPPVGAVTLSYPLSAVREERKASDGSVPGFGQLHPRTQGITTLGTIYSSS LFPGRAPEGHMLLLN YIGGTTNRGIVNQTTEQLVEQVDKDLRNMVIKPDAPKPRWGVRVWPRAIPQFNLGHLEQL DKARKALDAAGLQG VHLGGNYVSGVALGKWEHGYESAANLAKSVSKAAVKA

Uroporphyrinogen III decarboxylase (UROD1) nucleic acid sequence (SEQ ID NO: 41) atgcagaccaaggctttcacctctgcgcgcccccagcgggccgctgcgctcaaggcgcag cgcacctcgtcggtg accgtgcgcgcgaccgcggcccccgccgtggcctctgcccccgccgcctcgggctctgcc tctgaccccctgatg ctgcgcgccatccgcggcgacaaggtggagcgcccgcccgtgtggatgatgcgccaggcc ggccgctaccagaag gtgtaccaggacctgtgcaagaagcaccccacgttccgtgagcgctcggagcgcgtggac ctggcggtggagatc tctctgcagccgtggcacgcgttcaagcccgacggcgtcatcctgttcagcgacattctg acccccctgcccggc atgaacatccccttcgacatggcgcccggccccatcatcatggaccccatccgcaccatg gcgcaagtggagaag gtgacgaagctggacgctgaggccgcctgccccttcgtgggcgagtcgctgcgccagctg cgcacctacatcggc aaccaggccgcggtcctgggcttcgtgggcgcccccttcaccctggccacctacattgtg gagggcggcagctcc aagaacttcgcgcacatcaagaagatggctttctccacccccgagatcctgcacgccctg ctggacaagctggct gacaacgtggccgactacgtccgctaccaggccgacgccggcgcccaggtggtgcagatc ttcgactcgtgggcc agcgagctgcagccccaggacttcgacgtgttctccggcccctacatcaagaaggtgatc gacagcgtgcgcaag acccaccccgacctgcccatcatcctctacatcagcggctctggcggcctgctggagcgc atggcctcttgctcg cccgacatcatctcgctggaccagtcggtggacttcaccgacggcgtcaagcgctgcggc accaacttcgccttc cagggcaacatggaccccggcgtcctgttcggctccaaggacttcatcgagaagcgcgtc atggacaccatcaag gctgcccgcgacgccgacgtgcgccacgtgatgaacctgggccacggcgtgctgcccggc acccccgaggaccac gtgggccactacttccacgtcgcccgcaccgcccacgagcgcatgtaa

Uroporphyrinogen III decarboxylase (UROD1) amino acid sequence (SEQ ID NO: 42):

MQTKAFTSARPQRAAALKAQRTSSVTVRATAAPAVASAPAASGSASDPLMLRAIRGD KVERPPVWMMRQAGRYQK VYQDLCKKHPTFRERSERVDLAVEISLQPWHAFKPDGVILFSDILTPLPGMNIPFDMAPG PI IMDPIRTMAQVEK VTKLDAEAACPFVGESLRQLRTYIGNQAAVLGFVGAPFTLATYIVEGGSSKNFAHIKKMA FSTPEILHALLDKLA DNVADYVRYQADAGAQWQIFDSWASELQPQDFDVFSGPYIKKVIDSVRKTHPDLPIILYI SGSGGLLERMASCS PDIISLDQSVDFTDGVKRCGTNFAFQGNMDPGVLFGSKDFIEKRVMDTIKAARDADVRHV MNLGHGVLPGTPEDH VGHYFHVARTAHERM

Uroporphyrinogen III synthase (HEM4) nucleic acid sequence (SEQ ID NO: 43):

atgtcggccctggacgccgccgccatcccctacgagctagtgccgggtgtgtcctcc gctctggccgccccgctg ttcgccggcgtcccgctcacacacgtcagcctgagcccctcgttcaccgtggtcagcggg cacgacgtggccggc accgactgggcggcgttccgggggctgcccacgctggtggttctgatggcgggtcgtaac ctggggcagatagcc cggcggcttgtgcaggacgcggggtgggcgcccgatacacctgtaagtcaacctagtggc tag

Uroporphyrinogen III synthase (HEM4) amino acid sequence (SEQ ID NO: 44):

MSALDAAAJPYELVPGVSSALAAPLFAGVPLTHVSLSPSFTWSGHDVAGTDWAAFRGLPT LWLMAGRNLGQIA RRLVQDAGWAPDTPVSQPSG

CHUD 5’ untranslated region (regulatory region) (SEQ ID NO: 45):

ggcgtccccacaaccaggacagcctacttcttgaccttattaataagtcgctgcgtg tcgcgactgaccattttg gcccggacttgcgtgcttgtgatttgtgcttcgactagatccgcgggcaccaagggacgc ggacagctgatagtc aagaactagatcctctgggagcgtctggggctgtccccgctgctcgccaaggaa

CHUD 3’ untranslated region (regulatory region) (SEQ ID NO: 46):

gtgccgagtgactgaggtggcaaggtgcagtggcggcggaggcagttgtgctggggt ggcaaggcggacaggcga agctggtgggttgcgacgaggaggaggtgcacgtgcacgcgtaacataagaagaacagtg ggaggacaggtagcg tgacttgactgggacgaggagcgtactgatgtgtggcgtgtgttggtatgtgagcgttac ccctcccctagatag cggcggtctccactttcaggaggatgagagccatcatgaggctttgagggggcactggtt cgtgtgtaggctgag gctgctgttgaagtcacaaggcagcactgcatgcgcgagtgagtgtggccggatatgcat cgagttgcaggtaca ctgaaatgaggtgactgcggcgtatatcgctgccagtacaggttgaagcggcgggcacgg tgaatggagtactcg gcctggaacgcttgcgatcagatggtcgagctcaagaagatttggttgagccgttgggtc gtgcgtcatattatg gcttgcatcttcggggagcggcaagaaacggactccaatgcaggccctcgggcgagaaag attgggcgtgtccgg gggtgcattctcgccgcgtggggctgcatcgaatttcgcttgagtgccccttcccgggga gggggggcggtagtt caaccccatcatcgtaggggggttgtaaatgccagcccaaactaaa

CHUD Exon 1 (SEQ ID NO: 47) atgaagtctctctgccatgagctcgctggccccagcgttactgggtgcggccggcgaagc ctccggaaggctttc agcggtgccaagattgcgcaggtctctcgccccgctgtgcttaacagcgtgcagcgccaa cagcgtctcgcctgt tctgccgtggccgagctctccgctgctgagctgcgcg

CHLD Exon 2 (SEQ ID NO: 48):

ccatgaaggtgtctgaggaggactccaagggcttcgatgcggatgtgtcgacccgcc tggcccgctcgtaccctc tggcggccgtggtgggccaggacaacatcaagcaggcgctgctgctgggcgccgtggaca ccgggctgggcggca tcgccatcgccggtcgccgcggtaccgccaagtccatcatggctcgcggcctgcacgctc tgctgccgcccattg aggtggtggagggcagcatctgcaacgccgaccccgaggacccccgctcctgggag

CHLD Exon 3 (SEQ ID NO: 49):

gctggcctggctgagaagtatgcgggcggccctgtgaagaccaagatgcgctcggcg ccgtttgtgcagatccct ctgggtgtgactgaggaccgcttggtgggcactgtggacattgaggcgtccatgaag

CHLD exon 4 (SEQ ID NO: 50):

gagggcaagactgtgttccagcccggcctgctggctgaggcgcaccgcggcatcctg tacgtggacgagatcaac ctgctggatgacggcattgccaacctgctgctgtccatcctgtcggacggagtcaacgtg gtggagcgcgagggc atctccatcagccaccc

CHLD exon 5 (SEQ ID NO: 51):

ctgccggccgctgctgattgccacctacaaccccgaggagggccctctgcgtgagca cctgctggaccgcatcgc cattggcctcagcgccgacgtccccagcaccagcgacgagcgcgtcaaggc

cattgacgcagccatccgcttccaggacaagccgcag

CHLD exon 6 (SEQ ID NO: 52):

gacactattgacgacacc gcggagctcaccgacgccctgcgcacctcg

CHLD exon 7 (SEQ ID NO: 53):

gtcatcctggctcgcgagtacctgaaggacgtgaccatcgcgccggagcaggtgacc tacattgtggaggaggcg cgccgcggcggagtccaggggcacc gcgcggagctgtacgcggtcaag

CHLD exon 8 (SEQ ID NO: 54):

tgtgccaaggcgtgtgcggctctggagggccgtgagcgtgtgaacaaggatgacctg

cgccaggccgtgcagctggtcatcctgccgcgcgccaccatcctggaccagcccccg cccgagcaggagcagccc ccgccgccgcccccgccccctcccccgccgccgccgcag

CHLD exon 9 (SEQ ID NO: 55):

gaccaaatggaggacgaggaccaggaggagaaggaggacgagaaggaggaggaggag aaggagaacgaggaccag gacgagcccgag

CHLD exon 10 (SEQ ID NO: 56):

atccctcaggagttcatgtttgagtccgagggcgtcatcatggacccctccatcctc atgttcgcgcagcagcag cagcgcgcgcagggccgctccggccgcgccaagacgctcatcttcagcgacgaccgcggc cgctacatcaagccc atgctgcccaagggtgacaaggtcaagcgcctggcagtggacgccacgcttcgcgccgcc gcgcccta ccagaag

CHLD exon 11 (SEQ ID NO: 57) attcgccggcagcaggccatcagcgagggcaaggtgcagcgcaaggtgtacgtggacaag ccagaca

CHLD exon 12 (SEQ ID NO: 58):

tgcgctccaagaagctggcccgcaaggccggtgcgctggtgatttttgttgtggacg cgtccggctccatggctc tgaaccgcatgagcgccgccaagggcgcctgcatgcgcctgctggctgagtcgtacacca gccgcgaccaggtgt gcctcatccccttctacggcgacaaggccgaggtgctgctgccgccctccaagtccatcg ccatggcccgccgcc gcctggactcgctgccctgcggcggcggctcgccccttgcgcacggcctgtccacggcgg tacgtgtgggcatgc aggccagccaggcgggcgaggtgggccgcgtcatgatggtgctcatcacggacggccgcg ccaacgtcagcctgg ccaagtccaacgaggaccccgaggcgctcaagcccgacgcgcccaagcccaccgccgact cgctgaaggacgagg tgcgcgacatggccaagaaggccgcgtccgccggcatcaacgtgcttgtcattgacacgg agaacaagttcgtga gcaccggctttgcggaggagatctccaaggcagcgcagggcaagtactactacctgccca acgccagcgacgccg ccatcgcggcggccgcgtccggcgccatggccgcggccaagggcggctactag

CHLD Intron 1 (SEQ ID NO: 59):

gtgagcgcctactttgatatgtaccaaagataccactgataggtttaggcacggaag atctggacttggaccccg tttgcgcaagccgggcgatgcacccatttcgcggtcacgccgagcgctggggtgcaattt agcgtgcccgacaag ctagaaaacagggaattaccatttgtttaattttgttgcgagagatctttgcttgtgtcc accggccgcgcgggg gaacttccggtgttgcgcaaggttgcgtgcgtgcccaccatcaacacctgtgccaggtct gtgtcacccccaggt tccaccaccctgcaatcttccaattgtgtctcgtttgctcgttgtctaatagtcgtcctt tgctcatccctacct gcag

CHLD Intron 2 (SEQ ID NO: 60):

gtgaggcagggaaggtgacacaggaggttttgaaagagagacagggaggcaaagatg gatggcggggcgggcagt gactttggggcggcatggagtgggattggtggagtgggattgggcaccatgtatcacaga tgttggcaacacagc gcagggccttgctctgtgcttgtgttgaccgtctagtcccccgtgccctgaaccaagtct ttcctcctgacacgg tcctccatgtcctccttccggcattcccttcctcgtccacag

CHLD Intron 3 (SEQ ID NO: 61):

gtgagccagcaagggaggagaggggaacggccgggtagggcagccggagtttaacca cgccaattcaacggggag caacggggaagaggaagggccggaagaggacggcaaaagcatttggtgggggcagcggct gtagtcagaagcgca aaggctgccacagtgtggcccgcaccctcctcaccaccagtttggcatgatcgtttagca tgggctggaatactc accgccagttctctcctctcccctctcctcccctgtccccgcctgcag

CHLD Intron 4 (SEQ ID NO: 62):

gtgagtgcgcgcgctgggtgtgtttgtgggacggcgcggcattggagcgcaggtgcg ggtgctgggccgtgcact tgtccgttggttcccttggaagcttcgatacacactcttactgcacgctctttaaccgcc ccccccctccacctc tgcccgccccgtgcag

CHLD Intron 5 (SEQ ID NO: 63):

gtgggtgggggaaagtgactggatgtcggtgggttttaggtatgtgcgtgtgtacga tgcggggagcagtacgga agcgggcacgagcggtgagggggcaggattgtggcgcacgctcgggccaagcccgggctc gcgacagagggtggg cttgtattcgtagtcaagcgcatcaggaagtgcagttgactggattcacctgaaacggcg ctgagcgggcggcta atagaatcccgcttcctgtccgcccctccccttgcccttcaatccgtcag

CHLD Intron 6 (SEQ ID NO: 64):

gtgagtggcgggggccgtgcgtttgtttgttgcgtgggctggctggctggctttgtt ggatgagggcgctgctca ccactcatctctttgaatccccacttatccagttgcctgcatgaaaccccgcctgactca ctccccaccatcctg taccgcttttccaaacatccttgcaaccatcccgccatccccacccgcag

CHLD Intron 7 (SEQ ID NO: 65) gtgaggagttggagggggaaggggcgaggggatgcgacagaagcgagggcgaggggagcc ggggtgggttgttgc aagtgtcgtgaattatagaatgaccccaaaagcgccggcccaacagggcctattacttgc gagtcaatccaaccc ctgatatagggagaatggggtagaggtcgtatcacgacagcaaggatgtacagtgggcct tggggttgggaggta cagggaaaaaggagaggacatggggttgggtaagcggggaataacaaatatacacccagc gtttatggaagtggg agatggaaacgggggcggacgaacaggaacaggggccggatggaggggctatgggggcat ggtgggtgggggtac ggcgcggggcagagcagggtcttgggtgaatgggcaagatgctgatgcttgggatgaaga cactatgagcaaaga aatggttgttgacgattgccatgatcatcgcagtgggggaggcggggtggcaataccggc agtcaacagttgggg tgcgatcaagattgattggagtaccagcagtggccgggatctggctgacgtgtctcgagc gagttgctggggtgg caaggagatgcaggggcagacgacgttgtgcgaccacacttacacacatttccttcccct tgcgtgtgtccgtgc gccctgtgcctccag

CHLD Intron 8 (SEQ ID NO: 66):

gtacgtaaacgtatttgattgctcaggtggttagccttggtgtggctgctgtttgac ttgtgcagctgtctttgt gtacatgttccacaaccctgtactccccatattccgcccccattccag

CHLD Intron 9 (SEQ ID NO: 67):

gtgagaggcggcgcggcggcttgcgggcgaaggcggggggcggggcggaggcaatgc ggccgcgcatggccagca acggaagggctggctatcaacacggcgagcgcacgatattcatataagagtgccatcgtg caatgctgaatactt gcgccaaccggatctcgctgctccgcttccaccggactgctttctcatctctccccttca ccctgtgtgtatcca cag

CHLD Intron 10 (SEQ ID NO: 68):

gtgagtgcccgaggtggtgggtggtgaattggggcacgagggtatgtgggcctaagg gagctgaatggggcatgt tttcttctgagcatcacggtcagagcttgacctgtcctccccgctgtacccccgtgcacg gtccgacacag

CHLD Intron 11 (SEQ ID NO: 69):

gtgagtacagcgcatcccggcgcaatcattgggcctagttactgctgcaggactcgt gtgctcttaagggctggc agctgtcagaagctctactcctcgcactgaccactgtgcctttctctccttcctctctcc ctccccgcacccctc ctcccacttcctcaacag

CHLI2 5’ - untranslated region (regulatory region) (SEQ ID NO: 70):

gcagacttccataaagctcttgtaacgctgtaccaactagtaagcggtacaattcgc ctgagcccgagcaacgcg acctttcttgctctgtggatctctgataatctaaccagaccaaaaccttttcactaatct aggcaaca

CHLI2 3’ - untranslated region (regulatory region) (SEQ ID NO: 71):

aaaaggctggtgtaggcctgtcgggtcgtgttaaaggttgctgcgtgaacgtgtaag tgtgacagtgtgccggta tgtgtgtgtatacatgtgttgcggtgtgcttttgtggcggtacatggtgatgactgagcg ggtgggacagagcac ggttaactgacgagggcagtccgtgcgagacggacgtttttgtagccgaggtgcaaggac tgatgacgggctaag ctgctggagacttggagttgagagtgcaggtggatcgacggtttctctaaggagtatgaa taggcaggagggctg gagacatttggggtgcaaggaggcggtagtatgggagatgtccatgggcggattttggcc tctgtaacttcttaa cgccca

CHLI2 Exon 1 (SEQ ID NO: 72):

atgcagagtctccagggtcagcgcgcgttcactgcggtgcgccagggtcgggcgggt cccctgcggactcgcctg gtcgtgcgctcgtctgttgccttgccatccacgaaagccgcgaagaagccgaacttcccg ttcgtcaagattcag ggccaggaggagatgaagcttgcactgctgctgaacgtggtcgaccccaacatcggcgga gtgcttattatgggt gaccgcggcactgccaagtcggtcgcg

CHLI2 Exon 2 (SEQ ID NO: 73) gtccgcgccctggtggatatgcttcccgacattgacgtggttgagggcgacgccttcaac agctcccccaccgac cccaagttcatgggccccgacaccctgcagcgcttccgcaacggcgagaagctgcccacc gtccgcatgcggacc cccctg

CHLI2 Exon 3 (SEQ ID NO: 74):

gtggagctgcctctgggcgccaccgaggaccgcatctgcggcaccatcgacatcgag aaggcgctgacgcagggc atcaaggcctacgagcccggcctgctg

CHLI2 Exon 4 (SEQ ID NO: 75):

gccaaggccaaccgcggcatcctgtatgtggacgaggtgaacctgctggatgatggc ctg

CHLI2 Exon 5 (SEQ ID NO: 76):

gttgatgtcgtgctggactcgtcggctagcggcctgaacactgtggagcgtgagggt gtgtccattgtgcaccct gcccgcttcatcatgattggctcaggcaacccccag

CHLI2 Exon 6 (SEQ ID NO: 77):

gagggtgagctgcgcccgcagctgctggatcgcttcggcatgagcgtcaacgtggcc acgctgcaggacaccaag cagcgcacgcagctggtgctggaccg

CHLI2 Exon 7 (SEQ ID NO: 78):

gcttgcgtacgaggcggaccctgacgcatttgtggactcgtgcaaggccgagcagac ggcgctcacggacaagct ggaggcggcccgccagcgcctgcggtccgtcaagatcagcgaggagctgcag

CHLI2 Exon 8 (SEQ ID NO: 79):

atcctgatctcggacatttgctcgcgcctggatgtggatggcctgcgcggtgacatt gtgatcaaccgcgccgcc aaggcgcttgtggccttcgagggccgcaccgaggtgaccacgaatgacgtggagcgcgtc atctcgggctgcctc aaccaccg

CHLI2 Exon 9 (SEQ ID NO: 80):

cctgcgcaaggacccgctggaccccattgacaacggcaccaaggtggccatcctgtt caagcgcatgaccgaccc cgagatcatgaagcgcgaggaggaggccaagaagaagcgcgaggaggcggccgccaaggc caaggcggagggcaa ggcggaccgccccacgggcgccaaggctggcgcctgggctggcttgccccctcgtcggta a

CHLI2 Intron 1 (SEQ ID NO: 81):

gtaggtaacacaagcaattatggggcgaagatctaggctccgctgatccgggcgggc aatcggcatcgtcggtgc aaccgtggggcgtctgtgcaccctttgctggtgccaggttgcctgactcgcctgcattcc tgtaccgagccacat tggctgctttgcagcgtgcatgggacgggtgtaggataagcgctatgtatgcgatagcgc gggtgcaccggcttg gcatggcaaggttgcggggtgcacatgcgtgccagcgtcccctcagcatcagagtctgga tctaagggctcagcg gcttcctgcgcatgtgggtctttgcgtagtgctacgaagccttataattaaagctcatgt attgagtggtccggg tttggggcactagtagtgccaggaggcgcgtgccaggttgatatgagcatatcagcaccc gttccttgcgaaacg cttccgttgtgctcccttccccaccacctccccgctcatacccatacatatggctatccg tcctctcattgcttg cccctacag

CHLI2 Intron 2 (SEQ ID NO: 82):

gtgagcgggcctaccttctgaagacagtcttacgtgttgcactgcagcggtgttgcg cacctctgcttttgcgtg cgccgggaagcgcggattgcggcctcacagatcaagcccggaaacgcttgttgtttccag cgggtggcacacacg cgcgcgcgcgcacagtgacaccctcacggccgcgctgccctgcag CHLI2 Intron 3 (SEQ ID NO: 83)

gtgcgtagtgcatggggagaggggacgaggggaggagggcagggccaataaaccgaa ccccaagtcatcgagaca cagaacccgataatagctcccagatcgccaaggggtgaggcgggaagccaaggatgatgc gttggccgcattgcg tgttgacgtcaggcttacacagggtctgactggctgtgcttggggtttggcacgcttctt gactggccccgtacg catgctgcag

CHLI2 Intron 4 (SEQ ID NO: 84):

gtgagtggtggtggtttctgggtcagcagaggacttctgtagtaggtaatgtgggcc agggaagtgtggctaaca tgccaaacacgggggcgcaccagtgcaagctgcattcgctgacgtgcacgggtgcaatgg gtgcaaggcgaactg caatcgcggtgcacagttgccagggctgcgctcacgcttgagtgtctgcacacgcactgc ag

CHLI2 Intron 5 (SEQ ID NO: 85):

gtgcgtagcgtgcgcgcatgtacttgtctcccttgtcatgttgggaaaggtcggtcc ccagcctgcttgcaagat gcggccggtcagcagctgcggacggtcagcacctacgtgccgaggttgtgtaacatgaat ggcgttggggcggcc gacctgccacaagctgaactgcgaccagcaaggcagctgccagcaacgcacacccgacgt gctacacgcttgtgt tttgacctcctaaacacacccgcccgctgtctgtcacgtccacag

CHLI2 Intron 6 (SEQ ID NO: 86):

gtaagcggcggcggcgcggggacacggagggacatttcgcgagcatgggttgaggag tcgggaggattcggtggc tggccggagtcgggagtcggagtcgcgagtcggaagtcaagcttctggcggcttcgtgct gtcgggtgcgctcgc catgatggcgctgaccggagggcgtcacgctgtgtatgtgggcgcgcag

CHLI2 Intron 7 (SEQ ID NO: 87):

gtacggggcgtacagcgggggcggctgcacggggccagtgaccgacagggcagcacg cggctggcgaagagcgac aaagtgacagggtgaccaagaccgggtgatgccacgagaggggcgcgggagccgtgcatt gggtcgaggagggag gaatgcaactttacactgatgcctctgtatacggccgccttccgagccctgcaaaccttc gctttcccccgacgc acgcag

CHLI2 Intron 8 (SEQ ID NO: 88):

gtgagcgcagcgtgcggtggatgcggtgcgcgtgcgggttgccaacttattattttg tacgtggacgcgtggctg gcgatggcatgtcatggcgcgaatggatattgggcgaatggataccggtaatggtagcac ggggcggcagggcct ggcggtagtggggttgagggggcgaggactccagcgcgcgatacatgccatgttcagcat ggccccaactgacag cgcccgctgccctgtgcgccccgctccctccgcgcacccgctcctcctacacag

CHLH1 5’ - untranslated region (regulatory region) (SEQ ID NO: 89):

ctagtctagagggaactagggaggggcaacagagaa

CHLH1 3’ - untranslated region (regulatory region) (SEQ ID NO: 90):

gcggcctccccttcatggtagcactagttggcgggttgtggttggactaggcggcta gggtatatacctagtagc ggcggctgcggagtggagggctggcgcccagcgcgagggcgtggcctttcctcctggacc cgagagcgctccgcg aggagacggcgagtgagataggcagcagcgagcggagatcgatttgtgaacagttttgtg gcgggatcccatagc ggatgcagagaagaccttagagcagcttcctcggtggagtgaacgagccagagcggaggg aaggcgcatgaggga actgcagggactggaactgcgggagtgcaggtccggtgctaggtccgctaaacagtgcgg tctacgcctgtgtgt gaggtgtgcgtgtgtgtgtgagctgtgcggttttgttgtgcaaagtaggagtgagccgag ccgcgcgtactttgt ggcgtgtttggctgctggcgctgagagccaagagagggtaaacgggtttggtattttatg gtgcggggtgaaagc agccctcgcaggaatggagcgattctgcagcatgatgcacgtgtgcctgcgcgtggatgg tggctgttgatatgg ctctgccactccggcagcaccgctacgatacctagcggtgcctggagtggtctctctgtt tggtgcgtgatgttt gggtttgccgttttgattctttgtttcgtgctgaatggctgaggcggcaagacccctcgt gccagtgtacagagc ctcacggctccctcggaccccgcgtggggacgtccattcccggtggcggtgtcgcctcgg cggtgtaaagcaaaa aatatttt

CHLH1 Exon 1 (SEQ ID NO: 91):

atgcagacttcctcgcttcttggccggcgcacggcccacccggctgcgggcgcgacg cccaagccg

CHLH1 Exon 2 (SEQ ID NO: 92):

gttgcgccctcgccccgcgtggctagcacccgccag

CHLH1 Exon 3 (SEQ ID NO: 93):

gtcgcgtgcaatgtggcgactggaccccggccgcccatgaccaccttcaccggtggc aacaagggccctgctaag cagcaggtgtcgctggatctgcgcgacgagg

CHLH1 Exon 4 (SEQ ID NO: 94):

gcgctggcatgttcaccagcaccagcccggagatgcgccgtgtcgtccctgacgatg tgaagggtcgcgttaagg tgaaggttgtgtacgtggtgctggaggcccagtaccagtcggccatcagcgctgcggtga agaacatcaacgcca agaactccaag

CHLH1 Exon 5 (SEQ ID NO: 95):

gtgtgcttcgaggtggtgggctacctgctggaggagctgcgtgaccagaagaacctc gatatgctcaaggaggat gtggcctctgccaacatcttcatcggctcgctcatcttcattgaggagcttgccgagaag

CHLH1 Exon 6 (SEQ ID NO: 96):

attgtggaggcggtgagccccctgcgcgagaagctggacgcgtgcctgatcttcccg tccatgccggcggtcatg aagctgaacaagctgggcacgttttcgatggctcagctgggccagtcgaagtcggtgttc tcggagttcatcaag tctgctcgcaag

CHLH1 Exon 7 (SEQ ID NO: 97):

aacaacgacaacttcgaggagggcttgctgaagctggtgcgcaccctgcctaaggtg ctgaagtatctgccctcg gacaaggcgcaggacgccaagaacttcgtgaacagcctgcagtactggctgggcggtaac tcggacaacctggag aacctgctgctgaacaccgtcagcaactacgtgcccgctctgaagggcgtggacttcagc gtggctgagcccacc gcctaccccgatgtgggtatctggcaccctctggcctcgggcatgtacgaggacctgaag gagtacctgaactg

CHLH1 Exon 8 (SEQ ID NO: 98):

gtacgacacccgcaaggacatggtcttcgccaaggacgcccccgtcattggcctggt gctgcagcgctcgcacct ggtgactggcgatgagggccactacagcggcgtggtcgctgagctggagagccgcggtgc taaggtcatccccgt ctttgccg

CHLH1 Exon 9 (SEQ ID NO: 99):

gtggcctggacttctccgcccccgtcaagaagttcttctacgaccccctgggctctg gccgcacgttcgtggaca ccgttgtgtcgctgaccggcttcgcgctggtgggcggccccgcgcgccaggacgcgccga aggccattgaggcgc tgaagaacctgaacgtgccctacctggtgtcgctgccgctggtgttccagaccactgagg agtggctggacagcg agctgggcgtgcaccccgtccaggtggctctgcag

CHLH1 Exon 10 (SEQ ID NO: 100):

gttgccctgcccgagctggatggtgccatggagcccatcgtgttcgctggccgtgac tcgaacaccggcaagtcg cactcgctgcccgaccgcatcgcttcgctgtgcgctcgcgccgtgaactgggccaacctg cgcaagaagcgcaac gccgagaagaagctggccgtcaccgtgttcagcttcccccctgacaagggcaacgtcggc actgccgcctacctg aacgtgttcggctccatctaccgcgtgctgaagaacctgcagcgcgagggctacgacgtg ggcgccctgccgccc tcggaggaggatctgatccagtcggtgctgacccagaaggaggccaagttcaactcgacc gacctgcacatcgcc tacaagatgaaggtggacgagtaccagaagctgtgcccttacgccgaggcgctggaggag aactggggcaagccc cccggcaccctgaacaccaacggccaggagctgctggtgtacggccgccagtacggcaac gtcttcatcggcgtg cagcccaccttcggctacgagggcgacccgatgcgcctgctgttctcgaagtcggccagc ccccaccacggcttc gccgcctactacaccttcctggagaagatcttcaaggccgacgccgtgctgcacttcggc acccacggctcgctg gagttcatgcccggcaagcaggtcggcatgtcgggtgtgtgctaccccgactcgctgatc ggcaccatccccaac ctctactactacgccgccaacaacccgtctgaggccaccatcgccaagcgccgctcgtac gccaacaccattteg tacctgacgccgcctgccgagaacgccggcctgtacaagggcctgaaggagctgaaggag ctgatcagctcgtac cagggcatgcgtgagtctggccgcgccgagcagatctgcgccaccatcattgagaccgcc aagctgtgcaacctg gaccgcgacgtgaccctgcccgacgctgacgccaaggacctgaccatggacatgcgcgac agcgttgtgggccag gtgtaccgcaagctgatggagattgagtcccgcctgctgccctgcggcctgcacgtggtg ggctgcccgcccacc gccgaggaggccgtggccaccctggtcaacatcgctgagctggaccgcccggacaacaac ccccccatcaagggc atgcccggcatcctggcccgcgccattggtcgcgacatcgagtcgatttacagcggcaac aacaagggcgtcctg gctgacgttgaccagctgcagcgcatcaccgaggcctcccgcacctgcgtgcgcgagttc gtgaaggaccgcacc ggcctgaacggccgcatcggcaccaactggatcaccaacctgctcaagttcaccggcttc tacgtggacccctgg gtgcgcggcctgcagaacggcgagttcgccagcgccaaccgcgaggagctgatcaccctg ttcaactacctggag ttctgcctgacccag

CHLH1 Exon 11 (SEQ ID NO: 101):

gtggtcaaggacaacgagctgggcgccctggtagaggcgctgaacggccagtacgtc gagcccggccccggcggt gaccccatccgcaaccccaacgtgctgcccaccggcaagaacatccacgccctggaccct cagtcgattcccact caggccgcgctgaagagcgcccgcctggtggtggaccgcctgctggaccgcgagcgcgac aacaacggcggcaag taccccgagaccatcgcgctggtgctgtggggcactgacaacatcaagacctacggcgag tcgctggcccaggtc atgatgatggtcggtgtcaagcccgtggccgacgccctgggccgcgtgaacaagctggag gtgatccctctggag gagctgggccgcccccgcgtggacgtggttgtcaactgctcgggtgtgttccgcgacctg ttcgtgaaccagatg ctgctgctggaccgcgccatcaagctggcggccgagcaggacgagcccgatgagatgaac ttcgtgcgcaagcac gccaagcagcaggcggcggagctgggcctgcagagcctgcgcgacgcggccacccgtgtg ttctccaacagctcg ggctcctactcgtccaacgtcaacctggcggtggagaacagcagctggagcgacgagtcg cagctgcaggagatg tacctgaagcgcaagtcgtacgccttcaactcggaccg

CHLH1 Exon 12 (SEQ ID NO: 102):

ccccggcgccggtggcgagatgcagcgcgacgtgttcgagacggccatgaagaccgt ggacgtgaccttccagaa cctggactcgtccgagatctcgctgaccgatgtgtcgcactacttcgactccgaccccac caagctggtggcgtc gctgcgcaacgacggccgcacccccaacgcctacatcgccgacaccaccaccgccaacgc gcaggtccgcactct gggtgagaccgtgcgcctggacgcccgcaccaagctgctcaaccccaagtggtacgaggg catgcttgcctcggg ctacgagggcgtgcgcgagatccagaagcgcatgaccaacaccatgggctggtcggccac ctcgggcatggtgga caactgggtgtacgacgaggccaactcgaccttcatcgaggatgcggccatggccgagcg cctgatgaacaccaa ccccaacagcttccgcaagctggtggccaccttcctggaggccaacggccgcggctactg ggacgccaagcccga gcagctggagcgcctgcgccagctgtacatggacgtggaggacaagattgagggcgtcga ataa

CHLH1 Intron 1 (SEQ ID NO: 103):

gtaggtgtaattagaaggatcaaaacctagcggcctgatctgggactgacggcctcg cgcttcaatcactctgat gcag

CHLH1 Intron 2 (SEQ ID NO: 104):

gtaggcacggcagaatgctcaatgaacatgcagctacatatgtttgggatcatggct gatctctgtgcgacgggt ccgcgcag

CHLH1 Intron 3 (SEQ ID NO: 105):

gtgagcagcgcggaccgagcaagcgctggcgatgcagttggatttgttgttcttggg tcaggcgctcgctcgatg gccagcgcgtgtatttaatgggataagggttgagacaaagcatctcttcgggtaaaaatc ttagttttcgacagc acgttgagaggcatgcaacttgctctttcgcag CHLH1 Intron 4 (SEQ ID NO: 106)

gtgggtaaggagttgcattatcagtgtggcatggtgttgcgggcgtctggggcgctg caacagcggcatcgtgcc gaactgaccgtgccgggctacccgcgtgcag

CHLH1 Intron 5 (SEQ ID NO: 107):

gtgcgctagggttggggtctggagggtgtggattgcgcccaagtgccctgttgcgct tggcggtcgctgtcatga tgtgagggtgacgtagtgcactcaattgcctgctacgtcaccacctttgatgggctggat ctgaggcaggtcagc tcggttccctgctgcatccagtgtccctgtcgccctgcacgtttgacgctgttccccctt ccgcactgtctcgct ttgcag

CHLH1 Intron 6 (SEQ ID NO: 108):

gtgtgggcacgcgctttgggaagggaggcatacatttttggttgcggttaggctggg cgcggacttggcactcac acggtcattgcacactcatgtctcaccttcatttacggtcccttgtgccgaactacctac ag

CHLH1 Intron 7 (SEQ ID NO: 109):

gtgagcagcatcagggcagagtgcatgaacggattggtggcagtggggaatggaatt agacggacacgtctgggc ggcaatatgttgcgctgcagtttttggggtgtagtgaactagaaaatagggaagagatag gccacataacatccg aaagctcatatttttgcaaccggcgcacctatcacagcccacctgaagggttttgtagtc aacgcgtgcaactga ctagatgtccccttacctgtctgatttcag

CHLH1 Intron 8 (SEQ ID NO: 110):

gtgaggcggggcggcgctgccctcggtaggggttgcagatggtgatgggtaaccgaa tgcatggccaatggggag tgaaatcaggaaaggaggggtaacacaatgcagggcagcacctgaatcgtgaaggcggag ttaggcagggatctg tcagttcgcctgtcacgtggatgggcgcagctgacctttgtggtgttgtggtgtggcgca g

CHLH1 Intron 9 (SEQ ID NO: 111):

gtgagctcagctgggacatgtaggggctcgggtcgccggagcatcgatgtagaatta cgggaggaggggagaggg gagaggattgcacgaaccgagatgagggcggtggttcgggatttcgggcaaaagctcgtg cggcaagcgttcagt gactgaagagcagtgtgcttcaactgcccctctgtccctcag

CHLH1 Intron 10 (SEQ ID NO: 112):

gtgcgaccggtgccgctgcgtggccaacagcttggtgccaccttcctgcggtgttga tttacactgtgtgcgtgg atgtgttggtttttcgcaactttagtctgggctccagctctttgccttcattgatcactc gtcttacctcctgcg ccatcatttgaatacag

CHLH1 Intron 11 (SEQ ID NO: 113):

gtgagccttaatgcaacacgtgtagccgttcgcatgggtggctgggtcatgctatgg ttggatcggcgtccgcct gcttgctactgcctgttcggtaccagcgtttactgaccccgcgtgtgccattcccaccac ctaccccctcgcctt gcag

Ferrochelatase 5’ - Untranslated region (regulatory region) (SEQ ID NO: 114):

gacagtgatatagcaataccgatataataggtttggcgggcttcaccttgtccttac ccagaatgtggccctgac agtcgatttccagcccccttgccactcgctccctgatttcttcaatcaactagttgggtc gttttctcgtaagg

Ferrochelatase 3’ - Untranslated region (regulatory region) (SEQ ID NO: 115) gggggcgggtggcgagtaaggcgtatggcggagcgaggagatgggctgtggcgtggccgg tgttcttttgtgtga ttggaaacatagacggggtgcggcacgcggcctgactgctgcgcggttggtgtggttgcg gggggagcggggtcg atggggcagcgcgcacgagttggttgaaggaggagggccaggcgctgggctacacccatg gtttgaggatgctag tgagtgatgtgtgcggggggcatggtgtgtaccattcagagtccagatgcacgcacggtt gcgtgggagcgttcc ctgctgtgcatgatgatggcgccttcgatgaatcatctcttgaaggtccaaatgaaacgt ctgaagtctgcagag ggtggtgctggacatgccatccaggcggaagtgggcagctgtgtctgactacaaagtagg tcttgttttgcttgg atagcgtttggctatgtagcgtgtattctgctcatcaatcacgccaggcgtcagggacta cccatgcaagtcggg agcgtggctggctctggaaaagttgtagctgctaggtggcgttggctggggtgtcatgca tctcggcaggtaggc ggtagcggtggacgacctctgcagcggagcatgtgcacaagatgtgactgcgcatgcacc cgtatatgacggcgt tggcgtcagttgttgagagtgaacagaggagagacgagcgaagctgccatgcccttagtg gctggtgcgagaggg gaagaaagagagaggaaggactttgcggcagtgccccacgccggagttggggacacggtc atcaacagggcggcg gagctgggcggagtgggtgtgtgatgggacagggttcaaggcaggttggcgaggtcggag tgggtagaccagtcc ttcagtgcaagggcattagggcatgatgtaagggctgaagcttg

Ferrochelatase Exon 1 (SEQ ID NO: 116):

atggcgtcgtttggattgatgcaaaggacggtgcactgtccccagcttgtggaggag cggtgttcgccggtcgct ggctgctctggtcgtggcctgccagttatccagcggcaacg

Ferrochelatase Exon 2 (SEQ ID NO: 117):

gcgtggcgtgtgcagtgccaccaacggtgtccagcgagggcgtgtgctgcgccggac ggccgcttcgaccgacgt ggtctccttcgtggaccccaatgacattagaaaacccgcagcagcagcagctggccctgc ggtggataaggtcgg cgttctgctgttaaaccttggcgggcccgaaaagctcgacgacgtcaagcctttcctgta taacctattcgccga cccagaaattattcgcctgccagcggcagctcagttcctgcagccgctgctcgcgacgat catctccacgcttcg cgccccgaagagcgcggagggctatgaggccattggcggtggtagcccgttgcgtaggat tacagacgagcaggc ggaggcgctggcggagtctctgcgcgccaagggccaacctgcgaacgtgtacgtgggcat gcgctattggcaccc ctacacggaggaggcgctggagcacattaaggccgacggcgtcacgcgcctggtcatcct cccgctgtaccctca gttctccatctctaccagcggctccagccttcgactgcttgagtcgctcttcaagagcga catcgcgctcaagtc gctgcggcacacggtcatcccgtcctggtaccagcggcggggctacgtgagcgcgatggc ggacctgattgtaga

9

Ferrochelatase Exon 3 (SEQ ID NO: 118):

gagctgaagaagttccgggacgtgcccagcgtggagctgtttttctccgcgcacggc gtgcccaagtcctacgtg gaggaggcgggcgacccatacaaggaggagatggaggagtgcgtgcggctcattacggac gag

Ferrochelatase Exon 4 (SEQ ID NO: 119):

gtcaagcggcgcggcttcgccaacacgcacacgctggcctaccagagccgcgtgggc cccgcggaatggctcaag ccgtacacggatgagtccatcaa

Ferrochelatase Exon 5 (SEQ ID NO: 120):

ggagctgggcaagcgcggcgtcaagtcgctgctggcggtgcccatcagctttgtcag cgagcacattgagacgtt ggaggagatcgacatggagtaccgcgagctggcggaggagagcg

Ferrochelatase Exon 6 (SEQ ID NO: 121):

gcatccgcaactggggccgcgtgccggcgctgaacaccaacgccgccttcatcgacg acctggcggacgcggtga tggaggcgctgccctacgtgggctgcctggccgggccgacagactcgctggtgccgctgg

Ferrochelatase Exon 7 (SEQ ID NO: 122):

gcgacctggagatgctgctgcaggcctacgaccgcgagcgccgcacgctgccgtcac cggtggtgatgtgggagt ggggctggaccaagagcgcggagacgtggaacggccgcattgccatgattgccatcatca tcatcctggcgctgg aggcagccagcggccagtccatcctcaaaaacctgttcctggcggagtag Ferrochelatase Intron 1 (SEQ ID NO: 123)

gtgcgataataaatttgcatccttatgaattgctcaatgactaacgagcagcgtccg cgaccacag

Ferrochelatase Intron 2 (SEQ ID NO: 124):

gtgagggtggcattctgtaaagggagttgtggagttgggcagagcgagtgggtttgg tcgccagggcgaggatgt tgcgcgggcgttggcaggaacagggctgctagggcttgcgtggccagcgactagggtttc gactggccagcgccg ccggggcgcgcttgccgaagctgcacagccccaagcgcttctgtggatcaaatggaaact tgtggcagtgtgtat gctagcgccttggcgcaagaccaattttagtggtattactgttattactgtggtagcggt gggtattcggcggcg tggttgttgttgcagccccgtgcgactaagaccgctggcaacgacagcaagccgccgcac ccaggcatatacggc ccaccagcaccaccgtacacaaccacgtgcctttgcactctacgcaccacagcgcgctgc tgccgctcccacctc ccatcccaacggcccctcttacccccacttcacaacccctcctctcacacgccctcctct tccccctcctcttcc ag

Ferrochelatase Intron 3 (SEQ ID NO: 125):

gtgggccgggcgcagcgggcgggcgggaggggcaggaggggcaggaggggaggaagg gaggggaggaagggatgg aaagctggcgcagcggcagcggcgggacaggtagagggcgctgccccagcggcggcaggt gggcatggtgggcgg gtaggggcgacgcgtgagggactcgtcaggcatccgcatggcggcgacttgctgctcctc accgctgacggctgc atctgctgtgtgcgtaacctggcctggctggcaccgcag

Ferrochelatase Intron 4 (SEQ ID NO: 126):

gtgaggcccgtgggtgggacgcggggagggacgcggggagggggagacgcgggagcg ggacaagggtgaggatac ggggagggaataggagaggccatggggagggatggggacacgggaggatgcacgggcctg ggtggagccaggggg aagtggacgacgagcccggcgggaggagggctgggtagaaggacgcgggaggtggttggg acaggtggacggggc gtgtggagcatacggcgcaagaagcgggactgagcgggttgcagggatggatgtaatcac ggcaagtaagaaccc cgagtggggctcagcgtgtcagcctgccttatctttcgcgcaagcgctggggttttattt cgctgtacacacgtc gcgcctttctgccgcag

Ferrochelatase Intron 5 (SEQ ID NO: 127):

gtgaggaggcgccggagttttgggggaaggggtgcggcgtgaagcgagatggcaggg gcgaaggaaggagcggat ggtggctgggtgcaagcggagaggcgacagagagtggaggttttggtggagcggttgggg agaggggcgcagcag ggatgcggccctggggatggcgggacagaagggagcaagtttgccaagtgaagggggggg gtgctcaagaggaga gggcggtggaggttaagacggccgtgctggttatgctggggttgcaaggcgcatgggcgc atggagccgggggag tttggctgtggatgggcactgcggatgggcacggcttgctactcatgtgcggtcgcggtc cgcggtgtgtcagcc agccaggacccatcccactgggtcttcctgcgtgcctgggactgcttgccgccacccacc cattcatcaccacca ctgcgcagacccaccaacaccgctgccctgaactgctctgactcttggcgctcctcag

Ferrochelatase Intron 6 (SEQ ID NO: 128):

gtgagtcgcgccgtcgcggttggttcgcggatgccggttggcggatgacgttcggcg gttggcattgggtttggg tttgaggggttgttgggtgaggtcgggattggggtcgggattgggggtcgagcgtggggc tggcgtggatgatgg cgtggtctttggaaggggcttggggaggttgcgcgtgtggatgcggacagcatgggcgcg acagtgcgcatgtgc atgtgctgtgtcaaacgtctggtgcgttcagtgtgtccttgcgtgcctcccaccgtacgc agccatcccgcgcgc ctggaccgtagagaccgcctacgtgtccgctagcggcctcggcctcagcctaagcgccag tagcgccagcgacac aagcaacactgtcgctaatggcagcagcggcagcagcagcagtcacgagaatgcccgcgg ccgggagaaagtgct cctagccgggggccgccgctagctggtttcctcagcgcgtggacggtggtgccttcatcc cgaccaccccaggcg cgtccccagtcccgtcgagctcgcctgccttgtggcccgccttgaccgccctggcgccac ccggtggctcgcata acgactcgctttccgttctccgcctgacgctgtccgcctgacgctctgcgcttgactctt tgcgccttcctcccc tcttcccccag

Mutant sequenced RedAlgae CHLH DNA (SEQ ID NO: 129):

atgcagacttcctcgcttcttggccggcgcacggcccacccggctgcgggcgcgacg cccaagccggttgcgccc tcgccccgcgtggctagcacccgccaggtcgcgtgcaatgtggcgactggaccccggccg cccatgaccaccttc accggtggcaacaagggccctgctaagcagcaggtgtcgctggatctgcgcgacgagggc gctggcatgttcacc agcaccagcccggagatgcgccgtgtcgtccctgacgatgtgaagggtcgcgttaaggtg aaggttgtgtacgtg gtgctggaggcccagtaccagtcggccatcagcgctgcggtgaagaacatcaacgccaag aactccaaggtgtgc ttcgaggtggtgggctacctgctggaggagctgcgtgaccagaagaacctcgatatgctc aaggaggatgtggcc tctgccaacatcttcatcggctcgctcatcttcattgaggagcttgccgagaagattgtg gaggcggtgagcccc ctgcgcgagaagctggacgcgtgcctgatcttcccgtccatgccggcggtcatgaagctg aacaagctgggcacg ttttcgatggctcagctgggccagtcgaagtcggtgttctcggagttcatcaagtctgct cgcaagaacaacgac aacttcgaggagggcttgctgaagctggtgcgcaccctgcctaaggtgctgaagtatctg ccctcggacaaggcg caggacgccaagaacttcgtgaacagcctgcagtactggctgggcggtaactcggacaac ctggagaacctgctg ctgaacaccgtcagcaactacgtgcccgctctgaagggcgtggacttcagcgtggctgag cccaccgcctacccc gatgtgggtatctggcaccctctggcctcgggcatgtacgaggacctgaaggagtacctg aactggtacgacacc cgcaaggacatggtcttcgccaaggacgcccccgtcattggcctggtgctgcagcgctcg cacctggtgactggc gatgagggccactacagcggcgtggtcgctgagctggagagccgcggtgctaaggtcatc cccgtctttgccggt ggcctggacttctccgcccccgtcaagaagttcttctacgaccccctgggctctggccgc acgttcgtggacacc gttgtgtcgctgaccggcttcgcgctggtgggcggccccgcgcgccaggacgcgccgaag gccattgaggcgctg aagaacctgaacgtgccctacctggtgtcgctgccgctggtgttccagaccactgaggag tggctggacagcgag ctgggcgtgcaccccgtccaggtggctctgcaggttgccctgcccgagctggatggtgcc atggagcccatcgtg ttcgctggccgtgactcgaacaccggcaagtcgcactcgctgcccgaccgcatcgcttcg ctgtgcgctcgcgcc gtgaactgggccaacctgcgcaagaagcgcaacgccgagaagaagctggccgtcaccgtg ttcagcttcccccct gacaagggcaacgtcggcactgccgcctacctgaacgtgttcggctccatctaccgcgtg ctgaagaacctgcag cgcgagggctacgacgtgggcgccctgtccgccctcggaggaggatctgatccagtcggt gctgacccagaagga ggccaagttcaactcgaccgacctgcacatcgcctacaagatgaaggtggacgagtacca gaagctgtgccctta cgccgaggcgctggaggagaactggggcaagccccccggcaccctgaacaccaacggcca ggagctgctggtgta cggccgccagtacggcaacgtcttcatcggcgtgcagcccaccttcggctacgagggcga cccgatgcgcctgct gttctcgaagtcggccagcccccaccacggcttcgccgcctactacaccttcctggagaa gatcttcaaggccga cgccgtgctgcacttcggcacccacggctcgctggagttcatgcccggcaagcaggtcgg catgtcgggtgtgtg ctaccccgactcgctgatcggcaccatccccaacctctactactacgccgccaacaaccc gtctgaggccaccat cgccaagcgccgctcgtacgccaacaccatttcgtacctgacgccgcctgccgagaacgc cggcctgtacaaggg cctgaaggagctgaaggagctgatcagctcgtaccagggcatgcgtgagtctggccgcgc cgagcagatctgcgc caccatcattgagaccgccaagctgtgcaacctggaccgcgacgtgaccctgcccgacgc tgacgccaaggacct gaccatggacatgcgcgacagcgttgtgggccaggtgtaccgcaagctgatggagattga gtcccgcctgctgcc ctgcggcctgcacgtggtgggctgcccgcccaccgccgaggaggccgtggccaccctggt caacatcgctgagct ggaccgcccggacaacaacccccccatcaagggcatgcccggcatcctggcccgcgccat tggtcgcgacatcga gtcgatttacagcggcaacaacaagggcgtcctggctgacgttgaccagctgcagcgcat caccgaggcctcccg cacctgcgtgcgcgagttcgtgaaggaccgcaccggcctgaacggccgcatcggcaccaa ctggatcaccaacct gctcaagttcaccggcttctacgtggacccctgggtgcgcggcctgcagaacggcgagtt cgccagcgccaaccg cgaggagctgatcaccctgttcaactacctggagttctgcctgacccaggtggtcaagga caacgagctgggcgc cctggtagaggcgctgaacggccagtacgtcgagcccggccccggcggtgaccccatccg caaccccaacgtgct gcccaccggcaagaacatccacgccctggaccctcagtcgattcccactcaggccgcgct gaagagcgcccgcct ggtggtggaccgcctgctggaccgcgagcgcgacaacaacggcggcaagtaccccgagac catcgcgctggtgct gtggggcactgacaacatcaagacctacggcgagtcgctggcccaggtcatgatgatggt cggtgtcaagcccgt ggccgacgccctgggccgcgtgaacaagctggaggtgatccctctggaggagctgggccg cccccgcgtggacgt ggttgtcaactgctcgggtgtgttccgcgacctgttcgtgaaccagatgctgctgctgga ccgcgccatcaagct ggcggccgagcaggacgagcccgatgagatgaacttcgtgcgcaagcacgccaagcagca ggcggcggagctggg cctgcagagcctgcgcgacgcggccacccgtgtgttctccaacagctcgggctcctactc gtccaacgtcaacct ggcggtggagaacagcagctggagcgacgagtcgcagctgcaggagatgtacctgaagcg caagtcgtacgcctt caactcggaccgccccggcgccggtggcgagatgcagcgcgacgtgttcgagacggccat gaagaccgtggacgt gaccttccagaacctggactcgtccgagatctcgctgaccgatgtgtcgcactacttcga ctccgaccccaccaa gctggtggcgtcgctgcgcaacgacggccgcacccccaacgcctacatcgccgacaccac caccgccaacgcgca ggtccgcactctgggtgagaccgtgcgcctggacgcccgcaccaagctgctcaaccccaa gtggtacgagggcat gcttgcctcgggctacgagggcgtgcgcgagatccagaagcgcatgaccaacaccatggg ctggtcggccacctc gggcatggtggacaactgggtgtacgacgaggccaactcgaccttcatcgaggatgcggc catggccgagcgcct gatgaacaccaaccccaacagcttccgcaagctggtggccaccttcctggaggccaacgg ccgcggctactggga cgccaagcccgagcagctggagcgcctgcgccagctgtacatggacgtggaggacaagat tgagggcgtcgaata a

CHLI1 5’ - untranslated region (regulatory region) (SEQ ID NO: 130):

tcctacagagtaaaggtctaggcgatgcgcgactgaaagactgtgaatcccggcgtc gccgtggtgggatgtggg ccggtgcgctgtcgcagaggataaattacaggtatcaaacaaggttagggcgttggaagg agcggcgctagggaa ctgaaatcggatctgcatcggaccctcattccgcgacttgtccttcttttgcctcgcccc gcagctcttgagttt tgttcttgaccctttgacacgaaccaaccgatataaaa CHLI1 3’ - untranslated region (regulatory region) (SEQ ID NO: 131) gcggcaggccttcatggtcgtcgttggagcatttgcggaaaggctgatggcagcagatgc agccatgtcagttgt ggctgaagttgttggctggggcgggagcgggcagcagctgctgcgagcggccgaagcagc ggtgctgctttgcgt atgagaggaagaccagtgccctcgaggaggcgagtgcctgtgtgagtgtcaggacgtgtg acttcggaaactgag ggcggtgagtagatgtgactggggcttgcaggaagcctactgaccctatcagaaaaggtg agcaggggtatatgg tctaggagcgttgccggagcgtggctggccagtgctagccgcgcgggctctgttgctcgc tggcgcgccgccgcc ttcacaacagatgccgtagaaatgcagcgatgtgacgaggcgtggcctattctgcaatgt gtgaggcgccaatgg cgccactgacaaatggaggagtggtcaaagcttgggtacgttttgagagctgcatcgggc agcgaggatcagtgt gcggtaagaccgacggcagacggattggcaagggaataggagggacgtgggcgtgggcgc ccgcgctttgtcgag gccgcatgagccggccgcttctagacccgtagcccattttgaacaagcgcccacgcgtgc tcccgatgggggaca tcgatcacgggaattgattaaggggcatgtgtggtgtgcaagtgagtgactggtggttcc gtccctgtgaggttg tttcgttggacgtggctgccgggttgcgcgcgggctaagcgggcctgaggcagagcgctg gcgtgtagccgcgag tatcgatctgtaacgtgc

CHLI1 Exon 1 (SEQ ID NO: 132):

atggccctgaacatgcgtgtttcctcttccaaggtcgctgccaagcagcagggccgc atctccgcggtgccggtt gtgtcgagcaaggtggcctcctccgcccgcgtggcccccttccag

CHLI1 Exon 2 (SEQ ID NO: 133):

ggcgctcccgtggccgcgcagcgcgctgctctgctgg

CHLI1 Exon 3 (SEQ ID NO: 134):

tgcgcgccgctgccgctactgaggtcaaggctgctgagggccgcactgagaaggagc tgg

CHLI1 Exon 4 (SEQ ID NO: 135):

gccaggcccgccccatcttccccttcaccgccatcgtgggccaggatgagatgaagc tggcgctgattctgaacg tgatcgaccccaagatcggtggtgtcatgatcatgggcgaccgtggcactggcaagtcca ccaccattcgtgccc tggcggatctgctgcccgagatgcag

CHLI1 Exon 5 (SEQ ID NO: 136):

gtggttgccaacgacccctttaactcggaccccaccgaccccgagctgatgagcgag gaggtgcgcaaccgcgtc aaggccggcgagcagctgcccgtgtcttccaagaagattcccatggtggacctgcccctg ggcgccactgaggac cgcgtgtgcggcaccatcgacatcgagaaggcgctgaccgagg

CHLI1 Exon 6 (SEQ ID NO: 137):

gtgtcaaggcgttcgagcccggcctgctggccaaggccaaccgcggcatcctgtacg tggatgaggtcaacctgc tggacgaccacctg

CHLI1 Exon 7 (SEQ ID NO: 138):

gtcgatgtgctgctggactcggccgcctccggctggaacaccgtggagcgcgagggt atctccatcagccacccc gcccgcttcatcctggtcggctcgg

CHLI1 Exon 8 (SEQ ID NO: 139):

gcaaccccgaggagggtgagctgcgcccccagctgctggatcgcttcggcatgcacg cccagatcggcaccgtca aggacccccgcctgcgtgtgcagatcgtgtcgcagcgctcgaccttcgacgagaaccccg ccgccttccg

CHLI1 Exon 9 (SEQ ID NO: 140) caaggactacgaggccggccagatggcgctgacccagcgcatcgtggacgcgcgcaagct gctgaagcagggcga ggtcaactacgacttccgcgtcaagatcagccagatctgctcggacctgaacgtggacgg catccgcggcgacat cgtgaccaaccgcgccgccaaggccctggccgccttcgagggccgcaccgag

CHLI1 Exon 10 (SEQ ID NO: 141):

gtgacccccgaggacatctaccgtgtcattcccctgtgcctgcgccaccgcctccgg aaagaccccctggctgag atcgacgacggtgaccgcgtgcgtgagatcttcaagcaggtgttcggcatggagtaa

CHLI1 Intron 1 (SEQ ID NO: 142):

gtgtgcagttgcatctaaagaacgtccaattcatggttactgctcgtggatctaagc ggttggctcaccagcgtt ccatggtccccgattcgtgcacgcag

CHLI1 Intron 2 (SEQ ID NO: 143):

gtgagaagccatgatacaaatataaggatttgaagcggtagatctaggacccatcga acttgagcaccgacttgc agtccttgccttgtccggcgactgaacttctgcgcttgctttgcag

CHLI1 Intron 3 (SEQ ID NO: 144):

gtaagtgtcgcgcaaagattttctgccgggacgggtctccctcgcaacatctgaacc catggctcgtttttttgc cccgcag

CHLI1 Intron 4 (SEQ ID NO: 145):

gtgcgcgcctcccccaaccccagtttggcaaatgtgtggttaagcgtcgaaagcgtg aacagaaacaggtgttgc gggggccgcggaatggctgcaatgggtgctgggggcttcggagggtctgggggcgagttt gggtatacacgggcg cgcacacttgaaggaacgctcaaggacgacagcggaggcgtggagacagcgccggcccaa gcagcctgtacttgt agctgctggtcagctgaggcatcacgacttgggaccagcacccggcctcacggttgcaca aggccatcaccgcgc gccaccacccacgcctcttcaaacccatgccggcacctaccgctacccctgtgacacgct ccgcacacgccgccc cgcacaccccaccatgtgacag

CHLI1 Intron 5 (SEQ ID NO: 146):

gtgagagcgaggcgcggggcgtgctctgcaggctagggtgaagatcaggagagccga agcgggcccgaacagcgc agagagaggcaagacgacacccctgccgcgttttgatcacaagattcacacccttgctct ccccaacgctcccgc acatag

CHLI1 Intron 6 (SEQ ID NO: 147):

gtgagcaggggcagataggcggtcgggcggctgggcggcaggggctgtgttggctgt gttgggtgtgggctgagg ctggtgggtgggctggcgggtggcagggatagcggtgaggggatggtgatggggcagaat gggcgggtgggcgga cacgtggggtcgttgaagggtgtgtggggacggcaactggtatgcgatatgtcggcttgg ccctggcggggaaag cattcgcagaatggcgcacgaacgaggccggggagcgagcggggatgggagacgcaacct gcgctgcgaagtgcg gcgcgcgctccagttgacacgttgcacgaatgtggccagtgttcgcctgagagttatggg ttagaccgccagatg agccggttaagctggtggtcgcggttgatcggctgcttcccttccggttgcacgcctggc accctaacattaccc tgtccgctgctgccctttgcccacag

CHLI1 Intron 7 (SEQ ID NO: 148):

gtgagtgcagctgccgctgcggctgctgatggtgacctgtgcgaccacggggctccg catttctggacgaagcgt tgtaccatagccgtcttggtccctgatttgggccggctctggtccgaagccttgacatct acagttcaacatggc cgtataacgatcctgtgcccacccacacgccaccccgccag

CHLI1 Intron 8 (SEQ ID NO: 149) gtgagcgcgcgctctacgatacggcagacatgtacacactgcggcgcactgtagagcttg cattgcatttcaagg cctcgaaagagtagggtggtcgttctctggtggtgtccggccacaattatgcaccccggt gttggtgcagcagct gtgatgtcacaccttgcatcacccccctactgctgccgcctctcctctcttctcgcccgc ag

CHLI1 Intron 9 (SEQ ID NO: 150):

gtgagcagagcaatattgcagagggaagggtggcggaagggtgataacggttgggga tctagaggggcgagatgg atgcacacagcgcggggttggttatgcatgcctgcatggacgcgtgcacgcacccctgat ctgccggttttccaa ctggcgatgccgtattatgacctgcagctcaccatcctcatgcttgatttgcctcgctca g

CHLI1 Protein sequence (SEQ ID NO: 151):

MALNMRVSSSKVAAKQQGRISAVPWSSKVASSARVAPFQGAPVAAQRAALLVRAAAATEV KAAEGRTEKELGQA RPIFPFTAIVGQDEMKLALILNVIDPKIGGVMIMGDRGTGKSTTIRALADLLPEMQWAND PFNSDPTDPELMSE EVRNRVKAGEQLPVSSKKIPMVDLPLGATEDRVCGTIDIEKALTEGVKAFEPGLLAKANR GILYVDEVNLLDDHL VDVLLDSAASGWNTVEREGISISHPARFILVGSGNPEEGELRPQLLDRFGMHAQIGTVKD PRLRVQIVSQRSTFD ENPAAFRKDYEAGQMALTQRIVDARKLLKQGEVNYDFRVKISQICSDLNVDGIRGDIVTN RAAKALAAFEGRTEV TPEDIYRVIPLCLRHRLRKDPLAEIDDGDRVREIFKQVFGME

Mutant protein sequence RedAlgaeCHLH (SEQ ID NO: 152):

MQTSSLLGRRTAHPAAGATPKPVAPSPRVASTRQVACNVATGPRPPMTTFTGGNKGPAKQ QVSLDLRDEGAGMFT

STSPEMRRWPDDVKGRVKVKVVYWLEAQYQSAISAAVKNINAKNSKVCFEVVGYLLE ELRDQKNLDMLKEDVA

SANIFIGSLIFIEELAEKIVEAVSPLREKLDACLIFPSMPAVMKLNKLGTFSMAQLG QSKSVFSEFIKSARKNND

NFEEGLLKLVRTLPKVLKYLPSDKAQDAKNFVNSLQYWLGGNSDNLENLLLNTVSNY VPALKGVDFSVAEPTAYP

DVGIWHPLASGMYEDLKEYLNWYDTRKDMVFAKDAPVIGLVLQRSHLVTGDEGHYSG WAELESRGAKVIPVFAG

GLDFSAPVKKFFYDPLGSGRTFVDTWSLTGFALVGGPARQDAPKAIEALKNLNVPYL VSLPLVFQTTEEWLDSE

LGVHPVQVALQVALPELDGAMEPIVFAGRDSNTGKSHSLPDRIASLCARAVNWANLR KKRNAEKKLAVTVFSFPP

DKGNVGTAAYLNVFGSIYRVLKNLQREGYDVGALSALGGGSDPVGADPEGGQVQLDR PAHRLQDEGGRVPEAVPL

RRGAGGELGQAPRHPEHQRPGAAGVRPPVRQRLHRRAAHLRLRGRPDAPAVLEVGQP PPRLRRLLHLPGEDLQGR

RRAALRHPRLAGVHARQAGRHVGCVLPRLADRHHPQPLLLRRQQPV

CHLI1 DNA sequence (SEQ ID NO: 153):

atggccctgaacatgcgtgtttcctcttccaaggtcgctgccaagcagcagggccgc atctccgcggtgccggtt gtgtcgagcaaggtggcctcctccgcccgcgtggcccccttccagggcgctcccgtggcc gcgcagcgcgctgct ctgctggtgcgcgccgctgccgctactgaggtcaaggctgctgagggccgcactgagaag gagctgggccaggcc cgccccatcttccccttcaccgccatcgtgggccaggatgagatgaagctggcgctgatt ctgaacgtgatcgac cccaagatcggtggtgtcatgatcatgggcgaccgtggcactggcaagtccaccaccatt cgtgccctggcggat ctgctgcccgagatgcaggtggttgccaacgacccctttaactcggaccccaccgacccc gagctgatgagcgag gaggtgcgcaaccgcgtcaaggccggcgagcagctgcccgtgtcttccaagaagattccc atggtggacctgccc ctgggcgccactgaggaccgcgtgtgcggcaccatcgacatcgagaaggcgctgaccgag ggtgtcaaggcgttc gagcccggcctgctggccaaggccaaccgcggcatcctgtacgtggatgaggtcaacctg ctggacgaccacctg gtcgatgtgctgctggactcggccgcctccggctggaacaccgtggagcgcgagggtatc tccatcagccacccc gcccgcttcatcctggtcggctcgggcaaccccgaggagggtgagctgcgcccccagctg ctggatcgcttcggc atgcacgcccagatcggcaccgtcaaggacccccgcctgcgtgtgcagatcgtgtcgcag cgctcgaccttcgac gagaaccccgccgccttccgcaaggactacgaggccggccagatggcgctgacccagcgc atcgtggacgcgcgc aagctgctgaagcagggcgaggtcaactacgacttccgcgtcaagatcagccagatctgc tcggacctgaacgtg gacggcatccgcggcgacatcgtgaccaaccgcgccgccaaggccctggccgccttcgag ggccgcaccgaggtg acccccgaggacatctaccgtgtcattcccctgtgcctgcgccaccgcctccggaaagac cccctggctgagatc gacgacggtgaccgcgtgcgtgagatcttcaagcaggtgttcggcatggagtaa

[0134] Although the invention has been described with reference to the above example, it will be understood that modifications and variations are encompassed within the spirit and scope of the invention. Accordingly, the invention is limited only by the following claims.