Method and compositions for directed clonning and subclonning using homologous recombination

A technology of homology arm and recombinase, applied in the field of cloning

Inactive Publication Date: 2002-10-09
EURO LAB FUER MOLEKULARBIOLOGIE EMBL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Also, there is no literature showing that DNA cloning in E. coli sbcBC strains is possible

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  • Method and compositions for directed clonning and subclonning using homologous recombination
  • Method and compositions for directed clonning and subclonning using homologous recombination
  • Method and compositions for directed clonning and subclonning using homologous recombination

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Embodiment Construction

[0058] The present invention relates to a method and a composition for DNA cloning and subcloning using bacterial recombinase-mediated homologous recombination technology. The inventors of the present invention have found that bacterial recombinases can be used in a specific manner to obtain highly efficient target cloning or subcloning.

[0059]Bacterial recombinases preferably use RecE / T and / or Redα / β. In E. coli, the RecE / T pathway has been previously described and its components partially recognized (Hall and Kolodner, 1994, Proc. Natl. Acad. Sci. U.S.A. 91: 3205-3209; Gillen et al., 1981, J. Bacteriol. 145:521-532). Recombination through the RecE / T pathway requires the expression of two genes, rec E and rec T, the DNA sequences of which have been published (Hall et al., 1993, J. Bacteriol. 175:277-278). The function of the Rec E protein is similar to that of the λ exon, which is also called Redα, and the function of the Rec T protein is similar to that of the Redβ and e...

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Abstract

The present invention is directed to methods and compositions for DNA subcloning using bacterial recombinase-mediated homologous recombination. The invention relates to methods for cloning, compositions comprising polynucleotides usefall as cloning vectors, cells comprising such polynucleotide compositions, and kits useful for cloning mediated by bacterial recombinases, such as RecE / T and Redalpha / beta.

Description

[0001] 1 Overview [0002] The present invention relates to a method and a composition for DNA cloning and subcloning using bacterial recombinase-mediated homologous recombination technology. In a specific embodiment, bacterial homologous recombination is initiated using RecE / T or Redα / β recombinases, or any functionally equivalent system, such as erf from phage P22. In particular, the present invention relates to cloning methods, diagnostic methods, compositions containing polynucleotides useful as cloning vectors, cells containing said polynucleotide compositions, and cloning mediated by RecE / T and Redα / β Reagent test kit. 2. Background of the invention [0003] DNA cloning and subcloning in Escherichia coli is the basis of molecular biology. DNA cloning refers to a process in which an origin of replication is operably linked to a double-stranded DNA fragment and amplified in E. coli or other suitable host. DNA subcloning also refers ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/09C12N1/21C12N15/10C12N15/90C12Q1/68C12R1/19
CPCC12N15/10C12N15/902C12Q1/6897
Inventor F·A·斯图尔特Y·张J·P·P·穆伊雷尔斯
Owner EURO LAB FUER MOLEKULARBIOLOGIE EMBL
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