发明授权
US08852943B2 Method of modifying target region in host DNA and selectable marker cassette
有权
修改宿主DNA和选择性标记盒中靶区的方法
- 专利标题: Method of modifying target region in host DNA and selectable marker cassette
- 专利标题(中): 修改宿主DNA和选择性标记盒中靶区的方法
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申请号: US12933766申请日: 2009-03-23
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公开(公告)号: US08852943B2公开(公告)日: 2014-10-07
- 发明人: Katsutoshi Ara , Takuya Morimoto , Naotake Ogasawara
- 申请人: Katsutoshi Ara , Takuya Morimoto , Naotake Ogasawara
- 申请人地址: JP Tokyo
- 专利权人: Kao Corporation
- 当前专利权人: Kao Corporation
- 当前专利权人地址: JP Tokyo
- 代理机构: Sterne, Kessler, Goldstein & Fox P.L.L.C.
- 优先权: JP2008-076008 20080324; JP2009-044193 20090226
- 国际申请: PCT/JP2009/056409 WO 20090323
- 国际公布: WO2009/119862 WO 20091001
- 主分类号: C12N15/70
- IPC分类号: C12N15/70 ; C12N15/74 ; C12N15/10 ; C12N1/21 ; C07H21/04
摘要:
A method of modifying a target region in a host DNA using a donor DNA: wherein the donor DNA having regions homologous to a 5′-side region outside of the target region in the host DNA, a 3′-side region outside of the target region in the host DNA and a first homologous recombination region inside of the target region in the host DNA, respectively, in this order, and further having a first selectable marker gene, an expression-inducing promoter and a second selectable marker gene expressed under the control of the expression-inducing promoter between the region homologous to the 3′-side region and the region homologous to the first homologous recombination region; which method has the steps of: a first step of performing homologous recombination between the donor DNA and the host DNA at the regions of the 5′-side region and the first homologous recombination region, to conduct selection of a host integrated with the donor DNA based on expression of the first selectable marker gene; and a second step of performing homologous recombination, within the host DNA integrated with the donor DNA by the first step, between two regions of the 3′-side region derived from the host DNA and the 3′-side region derived from the donor DNA, to conduct selection of a host whose target region is modified based on expression of the second selectable marker gene under an expression-inducing condition for the expression-inducing promoter; and a selectable marker cassette for use in the method.