Title:
METHOD FOR CONSTRUCTING BIFUNCTIONAL REGULATORY ELEMENTS CAPABLE OF BOTH PROMOTION AND TERMINATION, AND LIBRARY OF BIFUNCTIONAL ELEMENTS
Document Type and Number:
WIPO Patent Application WO/2022/178741
Kind Code:
A1
Abstract:
The present invention relates to the technical field of synthetic biology. Disclosed are a method for constructing bifunctional regulatory elements capable of both promotion and termination, and a library of bifunctional elements. The construction method comprises: assembling a promoter sequence and a terminator sequence according to the principle of termination first and then promotion. The promoter sequence comprises an upstream activating sequence and a core promoter sequence, wherein the upstream activating sequence comprises Mig1/Mig2 binding sites, Rap1 binding sites, Gcr1 binding sites, weak Mig1 binding sites, a GC box, a CAAT box and an octamer box; and the core promoter sequence comprises a TATA box, a BRE element, an MTE element and an Inr element. The terminator sequence comprises an efficiency element, a position element and polyA. The bifunctional regulatory elements capable of both promotion and termination are used for pathway construction, thereby simplifying a construction process, with the pathway assembly efficiency being improved by approximately 18% compared with the prior art.
Inventors:
ZHANG GENLIN (CN)
LIU ZHENGYANG (CN)
NI XIAOXIA (CN)
ZHANG YAN (CN)
LIU ZHENGYANG (CN)
NI XIAOXIA (CN)
ZHANG YAN (CN)
Application Number:
PCT/CN2021/077808
Publication Date:
September 01, 2022
Filing Date:
February 25, 2021
Export Citation:
Assignee:
UNIV SHIHEZI (CN)
International Classes:
C12N15/81; C12N15/113; C12N15/65; C12P5/02
Foreign References:
| CN107326041A | 2017-11-07 | |||
| CN103898113A | 2014-07-02 |
Other References:
WANG ZHAOXIA: "Design of Terminator of Saccharomyces Cerevisiae and Application in Pathway Engineering", BASIC SCIENCES, CHINA MASTER’S THESES FULL-TEXT DATABASE, 15 January 2020 (2020-01-15), XP055961892
WEI LINNA: "Characterization and Application to Lycopene Synthesis Pathway of Terminators in Saccharomyces Cerevisiae", BASIC SCIENCES, CHINA MASTER’S THESES FULL-TEXT DATABASE, 15 January 2019 (2019-01-15), XP055961889
JIANG HUIHUI: "Research Progress of Promoter Elements in Pichia Pastoris Expression System", JOURNAL OF CHAOHU COLLEGE, vol. 19, no. 3, 31 December 2017 (2017-12-31), pages 67 - 72, XP055961888, ISSN: 1672-2868
TANG HONGTING, WU YANLING, DENG JILIANG, CHEN NANZHU, ZHENG ZHAOHUI, WEI YONGJUN, LUO XIAOZHOU, KEASLING JAY D.: "Promoter Architecture and Promoter Engineering in Saccharomyces cerevisiae", METABOLITES, vol. 10, no. 8, pages 320, XP055918931, DOI: 10.3390/metabo10080320
SARMA NAYAN J., KRISTINE WILLIS: "The new nucleoporin - Regulator of transcriptional repression and beyond ", NUCLEUS, vol. 3, no. 6, 9 October 2012 (2012-10-09), XP055961886, DOI: 10.4161/nucleus.22427
KEMLER IRIS, BUCHER ETIENNE, SEIPEL KATJA, MÜLLER-LMMERGLÜCK MICHAEL M., SCHAFFNER WALTER: "Promoters with the octamer DNA motif (ATGCAAAT) can be ubiquitous or cell type-specific depending on binding affinity of the octamer site and Oct-factor concentration", NUCLEIC ACIDS RESEARCH, OXFORD UNIVERSITY PRESS, GB, vol. 19, no. 2, 1 January 1991 (1991-01-01), GB , pages 237 - 242, XP055961882, ISSN: 0305-1048, DOI: 10.1093/nar/19.2.237
SANTANGELO GEORGE M.: "Glucose Signaling in Saccharomyces cerevisiae", MICROBIOLOGY AND MOLECULAR BIOLOGY REVIEWS, AMERICAN SOCIETY FOR MICROBIOLOGY, US, vol. 70, no. 1, 1 March 2006 (2006-03-01), US , pages 253 - 282, XP055961881, ISSN: 1092-2172, DOI: 10.1128/MMBR.70.1.253-282.2006
GERTZ JASON, BARAK A. COHEN: "Environment-specific combinatorial cis-regulation", MOLECULAR SYSTEMS BIOLOGY, THE MACMILLAN BUILDING, LONDON, GB, vol. 5, 1 January 2009 (2009-01-01), GB , XP055961880, ISSN: 1744-4292, DOI: 10.1038/msb.2009.1
WEI LINNA: "Characterization and Application to Lycopene Synthesis Pathway of Terminators in Saccharomyces Cerevisiae", BASIC SCIENCES, CHINA MASTER’S THESES FULL-TEXT DATABASE, 15 January 2019 (2019-01-15), XP055961889
JIANG HUIHUI: "Research Progress of Promoter Elements in Pichia Pastoris Expression System", JOURNAL OF CHAOHU COLLEGE, vol. 19, no. 3, 31 December 2017 (2017-12-31), pages 67 - 72, XP055961888, ISSN: 1672-2868
TANG HONGTING, WU YANLING, DENG JILIANG, CHEN NANZHU, ZHENG ZHAOHUI, WEI YONGJUN, LUO XIAOZHOU, KEASLING JAY D.: "Promoter Architecture and Promoter Engineering in Saccharomyces cerevisiae", METABOLITES, vol. 10, no. 8, pages 320, XP055918931, DOI: 10.3390/metabo10080320
SARMA NAYAN J., KRISTINE WILLIS: "The new nucleoporin - Regulator of transcriptional repression and beyond ", NUCLEUS, vol. 3, no. 6, 9 October 2012 (2012-10-09), XP055961886, DOI: 10.4161/nucleus.22427
KEMLER IRIS, BUCHER ETIENNE, SEIPEL KATJA, MÜLLER-LMMERGLÜCK MICHAEL M., SCHAFFNER WALTER: "Promoters with the octamer DNA motif (ATGCAAAT) can be ubiquitous or cell type-specific depending on binding affinity of the octamer site and Oct-factor concentration", NUCLEIC ACIDS RESEARCH, OXFORD UNIVERSITY PRESS, GB, vol. 19, no. 2, 1 January 1991 (1991-01-01), GB , pages 237 - 242, XP055961882, ISSN: 0305-1048, DOI: 10.1093/nar/19.2.237
SANTANGELO GEORGE M.: "Glucose Signaling in Saccharomyces cerevisiae", MICROBIOLOGY AND MOLECULAR BIOLOGY REVIEWS, AMERICAN SOCIETY FOR MICROBIOLOGY, US, vol. 70, no. 1, 1 March 2006 (2006-03-01), US , pages 253 - 282, XP055961881, ISSN: 1092-2172, DOI: 10.1128/MMBR.70.1.253-282.2006
GERTZ JASON, BARAK A. COHEN: "Environment-specific combinatorial cis-regulation", MOLECULAR SYSTEMS BIOLOGY, THE MACMILLAN BUILDING, LONDON, GB, vol. 5, 1 January 2009 (2009-01-01), GB , XP055961880, ISSN: 1744-4292, DOI: 10.1038/msb.2009.1
Attorney, Agent or Firm:
BEIJING GAOWO LAW FIRM (CN)
Download PDF: