发明申请
US20130023009A1 METHOD FOR SPECIFICALLY PRODUCING A JOINED DNA FRAGMENT COMPRISING A SEQUENCE DERIVED FROM A TARGET GENE
有权
特异性生产包含从目标基因衍生的序列的连接的DNA片段的方法
- 专利标题: METHOD FOR SPECIFICALLY PRODUCING A JOINED DNA FRAGMENT COMPRISING A SEQUENCE DERIVED FROM A TARGET GENE
- 专利标题(中): 特异性生产包含从目标基因衍生的序列的连接的DNA片段的方法
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申请号: US13394153申请日: 2010-09-02
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公开(公告)号: US20130023009A1公开(公告)日: 2013-01-24
- 发明人: Nobuyuki Kurosawa , Masaharu Isobe
- 申请人: Nobuyuki Kurosawa , Masaharu Isobe
- 申请人地址: JP Toyama-shi
- 专利权人: NATIONAL UNIVERSITY CORPORATION UNIVERSITY OF TOYAMA
- 当前专利权人: NATIONAL UNIVERSITY CORPORATION UNIVERSITY OF TOYAMA
- 当前专利权人地址: JP Toyama-shi
- 优先权: JP2009-205308 20090904
- 国际申请: PCT/JP10/64994 WO 20100902
- 主分类号: C12N15/66
- IPC分类号: C12N15/66 ; C12N15/11 ; C12N15/63 ; C12P19/34 ; C12P21/00
摘要:
Provided is a method for selectively obtaining, for a given target gene, a “joined DNA fragment” wherein just a target gene fragment is joined with desired other DNA fragments, regardless of whether a DNA fragment containing a target gene sequence has been purified. In the provided method, a double-stranded joining DNA fragment containing a sequence A and/or a sequence B is selectively joined to the ends of a target gene fragment. A mixture of a double-stranded gene fragment, the 3′ end of which is protruding, and the double-stranded joining DNA fragment, which are related in a prescribed manner, undergoes at least two cycles of thermal denaturation, reassociation, and DNA synthesis, resulting in a “joined DNA fragment,” which is a double-stranded DNA fragment including at least one instance of a sequence resulting from joining sequence A, the target gene sequence, and sequence B. A “single-side joined DNA fragment” can also be obtained, by a similar method.
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