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WO2009113794A3 - Methods for screening initial codons providing desired expression levels of proteins, and methods for tuning expression and production of recombinant proteins - Google Patents

Methods for screening initial codons providing desired expression levels of proteins, and methods for tuning expression and production of recombinant proteins Download PDF

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Publication number
WO2009113794A3
WO2009113794A3 PCT/KR2009/001183 KR2009001183W WO2009113794A3 WO 2009113794 A3 WO2009113794 A3 WO 2009113794A3 KR 2009001183 W KR2009001183 W KR 2009001183W WO 2009113794 A3 WO2009113794 A3 WO 2009113794A3
Authority
WO
WIPO (PCT)
Prior art keywords
methods
proteins
gene
production
expression
Prior art date
Application number
PCT/KR2009/001183
Other languages
French (fr)
Other versions
WO2009113794A2 (en
Inventor
Dong-Myung Kim
Jin-Ho Ahn
Jung-Won Keum
Original Assignee
The Industry & Academic Cooperation In Chungnam National University (Iac)
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by The Industry & Academic Cooperation In Chungnam National University (Iac) filed Critical The Industry & Academic Cooperation In Chungnam National University (Iac)
Publication of WO2009113794A2 publication Critical patent/WO2009113794A2/en
Publication of WO2009113794A3 publication Critical patent/WO2009113794A3/en

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/67—General methods for enhancing the expression
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/102—Mutagenizing nucleic acids
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/74—Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora

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  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Engineering & Computer Science (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biomedical Technology (AREA)
  • Organic Chemistry (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Biophysics (AREA)
  • Plant Pathology (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Abstract

Disclosed herein is a method for screening an initial codon providing a desired expression level of a protein by changing randomly the initial codon of the gene, introducing the gene into a cell to isolate an individual gene and performing a colony polymerase chain reaction (PCR) to amplify the gene, and expressing the gene to a protein using the cell-free protein synthesis system; and a method for controlling the expression of the recombinant protein; and a method for preparing the recombinant protein by using same.
PCT/KR2009/001183 2008-03-10 2009-03-10 Methods for screening initial codons providing desired expression levels of proteins, and methods for tuning expression and production of recombinant proteins WO2009113794A2 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR10-2008-0022217 2008-03-10
KR20080022217A KR100961565B1 (en) 2008-03-10 2008-03-10 Methods for screening initial codons providing desired expression levels of proteins, and methods for tuning expression and production of recombinant proteins

Publications (2)

Publication Number Publication Date
WO2009113794A2 WO2009113794A2 (en) 2009-09-17
WO2009113794A3 true WO2009113794A3 (en) 2009-12-23

Family

ID=41065660

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2009/001183 WO2009113794A2 (en) 2008-03-10 2009-03-10 Methods for screening initial codons providing desired expression levels of proteins, and methods for tuning expression and production of recombinant proteins

Country Status (2)

Country Link
KR (1) KR100961565B1 (en)
WO (1) WO2009113794A2 (en)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20130274129A1 (en) 2012-04-04 2013-10-17 Geneart Ag Tal-effector assembly platform, customized services, kits and assays
US11345959B2 (en) 2015-06-24 2022-05-31 Done Woo Lee Method for exploring useful genetic resources through bulk metagenome analysis and use thereof

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20040260060A1 (en) * 2000-06-22 2004-12-23 Laurent Chevalet Constructs modified downstreams of the initiation codon for recombinant protein
KR100733712B1 (en) * 2006-02-17 2007-06-29 충남대학교산학협력단 Preparation of cell extract and its application for cell-free protein systhesis

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20040260060A1 (en) * 2000-06-22 2004-12-23 Laurent Chevalet Constructs modified downstreams of the initiation codon for recombinant protein
KR100733712B1 (en) * 2006-02-17 2007-06-29 충남대학교산학협력단 Preparation of cell extract and its application for cell-free protein systhesis

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
GONZALEZ, V. E. ET AL.: "A codon window in mRNA downstream of the initiation codon where NGG codons give strongly reduced gene expression in Escherichia coli.", NUCLEIC ACIDS RES., vol. 32, no. 17, 2004, pages 5198 - 5205 *
QING, G. ET AL.: "Enhancement of translation initiation by AfT-rich sequences downstream of the initiation codon in Escherichia coli.", J. MOL. MICROBIOL. BIOTECHNOL., vol. 6, no. 3-4, 2003, pages 133 - 144 *
STENSTROM, C. M. ET AL.: "Influences on translation initiation and early elongation by the messenger RNA region flanking the initiation codon at the 3' side.'", GENE, vol. 288, no. 1-2, 17 April 2002 (2002-04-17), pages 1 - 8 *

Also Published As

Publication number Publication date
KR20090097056A (en) 2009-09-15
KR100961565B1 (en) 2010-06-07
WO2009113794A2 (en) 2009-09-17

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