发明申请
- 专利标题: In vitro recombination method
- 专利标题(中): 体外重组法
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申请号: US11502746申请日: 2006-08-11
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公开(公告)号: US20070037197A1公开(公告)日: 2007-02-15
- 发明人: Lei Young , Hamilton Smith , Daniel Gibson
- 申请人: Lei Young , Hamilton Smith , Daniel Gibson
- 主分类号: C12Q1/68
- IPC分类号: C12Q1/68 ; C12P19/34
摘要:
The present invention relates, e.g., to in vitro method, using isolated protein reagents, for joining two double stranded (ds) DNA molecules of interest, wherein the distal region of the first DNA molecule and the proximal region of the second DNA molecule share a region of sequence identity, comprising contacting the two DNA molecules in a reaction mixture with (a) a non-processive 5′ exonculease; (b) a single stranded DNA binding protein (SSB) which accelerates nucleic acid annealing; (c) a non strand-displacing DNA polymerase; and (d) a ligase, under conditions effective to join the two DNA molecules to form an intact double stranded DNA molecule, in which a single copy of the region of sequence identity is retained. The method allows the joining of a number of DNA fragments, in a predetermined order and orientation, without the use of restriction enzymes.
公开/授权文献
- US07723077B2 In vitro recombination method 公开/授权日:2010-05-25