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WO2003040318A3 - Method and system for inducible recombinational cloning in bacterial cells - Google Patents

Method and system for inducible recombinational cloning in bacterial cells Download PDF

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Publication number
WO2003040318A3
WO2003040318A3 PCT/US2002/035217 US0235217W WO03040318A3 WO 2003040318 A3 WO2003040318 A3 WO 2003040318A3 US 0235217 W US0235217 W US 0235217W WO 03040318 A3 WO03040318 A3 WO 03040318A3
Authority
WO
WIPO (PCT)
Prior art keywords
donor
plasmid
cassette
genes
transgene
Prior art date
Application number
PCT/US2002/035217
Other languages
French (fr)
Other versions
WO2003040318A2 (en
Inventor
Patrick Y Lu
Qingquan Tang
Martin C Woodle
Original Assignee
Intradigm Corp
Patrick Y Lu
Qingquan Tang
Martin C Woodle
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 Intradigm Corp, Patrick Y Lu, Qingquan Tang, Martin C Woodle filed Critical Intradigm Corp
Priority to AU2002356891A priority Critical patent/AU2002356891A1/en
Publication of WO2003040318A2 publication Critical patent/WO2003040318A2/en
Publication of WO2003040318A3 publication Critical patent/WO2003040318A3/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
    • C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
    • C12N15/90—Stable introduction of foreign DNA into chromosome
    • C12N15/902—Stable introduction of foreign DNA into chromosome using homologous recombination
    • 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
    • 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
    • C12N15/64—General methods for preparing the vector, for introducing it into the cell or for selecting the vector-containing host

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  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
  • Organic Chemistry (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Zoology (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • Physics & Mathematics (AREA)
  • Plant Pathology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Biophysics (AREA)
  • Cell Biology (AREA)
  • Mycology (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)

Abstract

The present invention discloses methods and systems that enable high throughput screening of genes and DNA sequence bearing expressed sequence tags in mammalian cells. The invention can easily be applied to the delivery of other functions like that of RNAi, ribozyme, antisense, DNA vaccine, etc.. The invention comprises a donor plasmid bearing a donor cassette and a receiver plasmid bearing a recombinase cassette, which may be propagated and maintained in bacterial culture. Through the use of inducible recombinational cloning, the donor cassette comprising a transgene sequence and a receiver plasmid, are easily and efficiently recombined in vivo to create a target vector comprising desired motifs (e.g. splice sites, introns and polyadenylation sequences). The present invention also provides convenient means for the introduction of the transgene into the donor cassette and provides an ability to create a library of transgene sequences for study maintained either in the donor plasmid or in the target vector. Because of the ease with which genes and putative genes may be introduced into the donor plasmid, and that recombination occurs in vivo, the invention allows for the functional study of genomics in model systems.
PCT/US2002/035217 2001-11-02 2002-11-04 Method and system for inducible recombinational cloning in bacterial cells WO2003040318A2 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU2002356891A AU2002356891A1 (en) 2001-11-02 2002-11-04 Method and system for inducible recombinational cloning in bacterial cells

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US33091001P 2001-11-02 2001-11-02
US60/330,910 2001-11-02

Publications (2)

Publication Number Publication Date
WO2003040318A2 WO2003040318A2 (en) 2003-05-15
WO2003040318A3 true WO2003040318A3 (en) 2003-11-06

Family

ID=23291829

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2002/035217 WO2003040318A2 (en) 2001-11-02 2002-11-04 Method and system for inducible recombinational cloning in bacterial cells

Country Status (2)

Country Link
AU (1) AU2002356891A1 (en)
WO (1) WO2003040318A2 (en)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101845445B (en) * 2010-02-04 2012-05-09 东北农业大学 Cre recombinase gene and Cre/loxP-mediated transgenic safety plant expression vector
CN108642086B (en) * 2018-05-14 2022-06-07 北京工业大学 A highly sensitive fusion cell screening method

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1997047758A1 (en) * 1996-06-14 1997-12-18 Massachusetts Institute Of Technology Methods and compositions for transforming cells
WO2000012687A1 (en) * 1998-08-28 2000-03-09 Invitrogen Corporation System for the rapid manipulation of nucleic acid sequences
US6274354B1 (en) * 1996-09-06 2001-08-14 The Trustees Of The University Of Pennsylvania Methods using cre-lox for production of recombinant adeno-associated viruses
US6277608B1 (en) * 1997-10-24 2001-08-21 Invitrogen Corporation Recombinational cloning using nucleic acids having recombination sites
US6303327B1 (en) * 1996-12-06 2001-10-16 Harald Von Melchner Gene trap construct for identification and isolation of genes

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1997047758A1 (en) * 1996-06-14 1997-12-18 Massachusetts Institute Of Technology Methods and compositions for transforming cells
US6274354B1 (en) * 1996-09-06 2001-08-14 The Trustees Of The University Of Pennsylvania Methods using cre-lox for production of recombinant adeno-associated viruses
US6303327B1 (en) * 1996-12-06 2001-10-16 Harald Von Melchner Gene trap construct for identification and isolation of genes
US6277608B1 (en) * 1997-10-24 2001-08-21 Invitrogen Corporation Recombinational cloning using nucleic acids having recombination sites
WO2000012687A1 (en) * 1998-08-28 2000-03-09 Invitrogen Corporation System for the rapid manipulation of nucleic acid sequences

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
DAVIES ET AL.: "An antibody VH domain with a lox-Cre site integrated into its coding region: Bacterial recombination with a single polypeptide chain", FEBS LETTERS, vol. 377, 1995, pages 92 - 96, XP000891732 *

Also Published As

Publication number Publication date
WO2003040318A2 (en) 2003-05-15
AU2002356891A1 (en) 2003-05-19

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