WO2005075644A3 - Leader sequence and method for synthesizing polypepides in cell-free systems - Google Patents

Leader sequence and method for synthesizing polypepides in cell-free systems Download PDF

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Publication number
WO2005075644A3
WO2005075644A3 PCT/EP2005/001146 EP2005001146W WO2005075644A3 WO 2005075644 A3 WO2005075644 A3 WO 2005075644A3 EP 2005001146 W EP2005001146 W EP 2005001146W WO 2005075644 A3 WO2005075644 A3 WO 2005075644A3
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WO
WIPO (PCT)
Prior art keywords
leader sequence
cell
poly
free systems
mrna
Prior art date
Application number
PCT/EP2005/001146
Other languages
French (fr)
Other versions
WO2005075644A2 (en
Inventor
Anatoly Timofeevich Gudkov
Marija Vladimirovna Ozerova
Vyacheslav Mihailovich Shiryaev
Alexander-Sergeyevich Spirin
Original Assignee
Roche Diagnostics Gmbh
Hoffmann La Roche
Inst Of Protein Res
Anatoly Timofeevich Gudkov
Marija Vladimirovna Ozerova
Shiryaev Vyacheslav Mihailovic
Alexander-Sergeyevich Spirin
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 Roche Diagnostics Gmbh, Hoffmann La Roche, Inst Of Protein Res, Anatoly Timofeevich Gudkov, Marija Vladimirovna Ozerova, Shiryaev Vyacheslav Mihailovic, Alexander-Sergeyevich Spirin filed Critical Roche Diagnostics Gmbh
Publication of WO2005075644A2 publication Critical patent/WO2005075644A2/en
Publication of WO2005075644A3 publication Critical patent/WO2005075644A3/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
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P21/00—Preparation of peptides or proteins
    • C12P21/02—Preparation of peptides or proteins having a known sequence of two or more amino acids, e.g. glutathione
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/60—Fusion polypeptide containing spectroscopic/fluorescent detection, e.g. green fluorescent protein [GFP]

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  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Plant Pathology (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Enzymes And Modification Thereof (AREA)

Abstract

The invention pertains to the leader sequence that provides for enhancement of mRNA translation in cell-free systems. The 5’-UTR leader sequence contains poly(A) region joined to the initiator mRNA codon of up to 25A long or a deletion of the poly(A) region in the range from 25A to 5A, or a combination of the poly(A) region with at least one nucleotide, or a substitution of at least one nucleotide in a complete or deleted form of poly(A). The invention pertains also to the method for synthesis of polypeptides in cell-free systems, wherein at the first stage a genetic construct is obtained that contains a leader sequence in the 5’-UTR which includes poly(A); the length and composition of the leader sequence are chosen by the results of mRNA translation or coupled transcription-translation of DNA in eukaryotic cell-free systems at mRNA concentrations from 50 to 2000 pmol/ml.
PCT/EP2005/001146 2004-02-06 2005-02-04 Leader sequence and method for synthesizing polypepides in cell-free systems WO2005075644A2 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
RU2004103495 2004-02-06
RU2004103495/13A RU2004103495A (en) 2004-02-06 2004-02-06 LEADER SEQUENCE AND METHOD FOR SYNTHESIS OF POLYEPEPTIDES IN A CELLLESS SYSTEM

Publications (2)

Publication Number Publication Date
WO2005075644A2 WO2005075644A2 (en) 2005-08-18
WO2005075644A3 true WO2005075644A3 (en) 2008-12-11

Family

ID=34836927

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2005/001146 WO2005075644A2 (en) 2004-02-06 2005-02-04 Leader sequence and method for synthesizing polypepides in cell-free systems

Country Status (2)

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RU (1) RU2004103495A (en)
WO (1) WO2005075644A2 (en)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ES2946969T3 (en) * 2014-12-12 2023-07-28 CureVac SE Artificial nucleic acid molecules for enhanced protein expression

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2385134A1 (en) * 1999-10-15 2001-04-19 Wakenyaku Co., Ltd. Template molecule having broad applicability and highly efficient function means of cell-free synthesis of proteins by using the same

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2385134A1 (en) * 1999-10-15 2001-04-19 Wakenyaku Co., Ltd. Template molecule having broad applicability and highly efficient function means of cell-free synthesis of proteins by using the same

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
AHN B-Y ET AL: "IDENTIFICATION OF THE VACCINIA VIRUS GENE ENCODING AN 18-KILODALTON SUBUNIT OF RNA POLYMERASE AND DEMONSTRATION OF A 5' POLY-A LEADER ON ITS EARLY TRANSCRIPT", JOURNAL OF VIROLOGY, vol. 64, no. 6, 1990, pages 3019 - 3024, XP002341120, ISSN: 0022-538X *
DE MELO NETO OSVALDO P ET AL: "Autoregulation of poly(A)-binding protein synthesis in vitro", NUCLEIC ACIDS RESEARCH, vol. 23, no. 12, 1995, pages 2198 - 2205, XP002341118, ISSN: 0305-1048 *
GUDKOV ANATOLY T ET AL: "5'-poly(A) sequence as an effective leader for translation in eukaryotic cell-free systems.", BIOTECHNOLOGY AND BIOENGINEERING. 20 AUG 2005, vol. 91, no. 4, 20 August 2005 (2005-08-20), pages 468 - 473, XP002341121, ISSN: 0006-3592 *
PATEL D D ET AL: "A POXVIRUS-DERIVED VECTOR THAT DIRECTS HIGH LEVELS OF EXPRESSION OF CLONED GENES IN MAMMALIAN CELLS", PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF USA, NATIONAL ACADEMY OF SCIENCE. WASHINGTON, US, vol. 85, no. 24, 1 December 1988 (1988-12-01), pages 9431 - 9435, XP000008759, ISSN: 0027-8424 *
PATEL D D ET AL: "Messenger RNAs of a strongly-expressed late gene of cowpox virus contain 5'-terminal poly(A) sequences", EMBO (EUROPEAN MOLECULAR BIOLOGY ORGANIZATION) JOURNAL, vol. 6, no. 12, 1987, pages 3787 - 3794, XP002341119, ISSN: 0261-4189 *

Also Published As

Publication number Publication date
RU2004103495A (en) 2005-07-27
WO2005075644A2 (en) 2005-08-18

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