Title:
HETEROLOGOUS EXPRESSION OF FUNGAL POLYKETIDE SYNTHETIC GENE IN YEAST
Document Type and Number:
WIPO Patent Application WO/2012/020574
Kind Code:
A1
Abstract:
Provided is a method which comprises, from a gene containing an eukaryotic intron, removing the intron contained in the gene and ligating exon sequences alone to give an expression vector carrying the thus ligated sequences. Specifically provided are: a method which comprises, from a giant fungal gene containing an intron, amplifying exon sequences, as one or more fragments, by PCR, and ligating the fragments to a vector, which has been treated with a restriction enzyme, by using the gap-repair-cloning method to thereby give an expression vector containing the ligated exon sequences; a method which comprises synthesizing cDNA fragments of a giant gene and ligating the fragments to give an expression vector containing a full-length cDNA sequence; a transformant having an expression vector, which is produced by the aforesaid method, transferred therein; a protein produced by the aforesaid transformant; and a method for obtaining a compound, which is produced by the aforesaid protein, using the aforesaid expression vector.
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Inventors:
WATANABE KENJI (JP)
MORIYA HISAO (JP)
MORIYA HISAO (JP)
Application Number:
PCT/JP2011/004566
Publication Date:
February 16, 2012
Filing Date:
August 11, 2011
Export Citation:
Assignee:
SHIZUOKA PREFECTURE PUBLIC UNIVERSITY CORP (JP)
UNIV OKAYAMA NAT UNIV CORP (JP)
WATANABE KENJI (JP)
MORIYA HISAO (JP)
UNIV OKAYAMA NAT UNIV CORP (JP)
WATANABE KENJI (JP)
MORIYA HISAO (JP)
International Classes:
C12N15/09; C12N1/15; C12N1/19; C12N1/21; C12N5/10
Domestic Patent References:
| WO1999066035A2 | 1999-12-23 |
Foreign References:
| JP2010181279A | 2010-08-19 | |||
| JP2011007312A | 2011-01-13 |
Other References:
DATABASE GENBANK [online] 9 April 2008 (2008-04-09), "Chaetomium globosum CBS 148.51 hypothetical protein (CHGG_10128) partial mRNA.", XP003033102, accession no. NCBI Database accession no. XM_001228054
CHINO A. ET AL.: "Plasmid construction using recombination activity in the fission yeast Schizosaccharomyces pombe", PLOS ONE, vol. 5, no. 3, March 2010 (2010-03-01), pages E9652-1 - E9652-7, XP055095577
MATSUO Y. ET AL.: "Simple and effective gap-repair cloning using short tracts of flanking homology in fission yeast", BIOSCI. BIOTECHNOL. BIOCHEM., vol. 74, no. 3, March 2010 (2010-03-01), pages 685 - 689, XP055095579
TAKAYOSHI SARUWATARI ET AL.: "Spliceosome Hi Izon-gata Genome DNA no Hatsugen ni yoru Polyketide-kei Tennenbutsu no Shinki Tansakuho", SYMPOSIUM ON BIOCATALYST CHEMISTRY JAPAN KOEN YOSHISHU, vol. 14, September 2010 (2010-09-01), pages 26 - 27
NORIYASU ISHIKAWA ET AL.: "Spliceosome Hi Izon-gata Genome DNA no Hatsugen ni yoru Polyketide-kei Tennenbutsu no Shinki Tansakuho", ABSTRACTS OF ANNUAL MEETING OF PHARMACEUTICAL SOCIETY OF JAPAN, vol. 131, no. 2, March 2011 (2011-03-01), pages 74
LEADLAY, P. ET AL., NATURE, 1990
KATZ, L. ET AL., SCIENCE, 1991
SAMSON, S. ET AL., NATURE, 1985
HISAO MORIYA ET AL., PLOS ONE, 2010
JAY D. KEASLING ET AL., NATURE, 2006
CHINO A. ET AL.: "Plasmid construction using recombination activity in the fission yeast Schizosaccharomyces pombe", PLOS ONE, vol. 5, no. 3, March 2010 (2010-03-01), pages E9652-1 - E9652-7, XP055095577
MATSUO Y. ET AL.: "Simple and effective gap-repair cloning using short tracts of flanking homology in fission yeast", BIOSCI. BIOTECHNOL. BIOCHEM., vol. 74, no. 3, March 2010 (2010-03-01), pages 685 - 689, XP055095579
TAKAYOSHI SARUWATARI ET AL.: "Spliceosome Hi Izon-gata Genome DNA no Hatsugen ni yoru Polyketide-kei Tennenbutsu no Shinki Tansakuho", SYMPOSIUM ON BIOCATALYST CHEMISTRY JAPAN KOEN YOSHISHU, vol. 14, September 2010 (2010-09-01), pages 26 - 27
NORIYASU ISHIKAWA ET AL.: "Spliceosome Hi Izon-gata Genome DNA no Hatsugen ni yoru Polyketide-kei Tennenbutsu no Shinki Tansakuho", ABSTRACTS OF ANNUAL MEETING OF PHARMACEUTICAL SOCIETY OF JAPAN, vol. 131, no. 2, March 2011 (2011-03-01), pages 74
LEADLAY, P. ET AL., NATURE, 1990
KATZ, L. ET AL., SCIENCE, 1991
SAMSON, S. ET AL., NATURE, 1985
HISAO MORIYA ET AL., PLOS ONE, 2010
JAY D. KEASLING ET AL., NATURE, 2006
Attorney, Agent or Firm:
OHNO, Seiji et al. (JP)
Seiji Ono (JP)
Seiji Ono (JP)
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