Method for constructing and modifying large DNA molecules
    1.
    发明授权
    Method for constructing and modifying large DNA molecules 有权
    构建和修饰大DNA分子的方法

    公开(公告)号:US07781190B2

    公开(公告)日:2010-08-24

    申请号:US10892579

    申请日:2004-07-16

    IPC分类号: C12N15/64 C12N15/70

    CPC分类号: C12N15/10

    摘要: This invention provides a method for combining overlapping DNA molecules comprising: (a) providing first and second DNA fragments, the first having a region homologous to a region in the second; (b) tagging the first DNA fragment with a selectable marker; (c) cloning the first DNA sequence into a retrieval vector to form a DNA-vector complex; (d) linearizing the DNA-vector complex; and (e) inserting the first DNA fragment from the DNA-vector complex into the second DNA fragment using homologous recombination to form a combined DNA molecule; and (f) removing the selectable marker, thereby generating a combined DNA molecule. The invention further provides a vector for retrieving and inserting a selected DNA molecule into a target DNA molecule.

    摘要翻译: 本发明提供了一种组合重叠DNA分子的方法,包括:(a)提供第一和第二DNA片段,第一DNA片段具有与第二DNA片段中的区域同源的区域; (b)用选择标记标记第一个DNA片段; (c)将第一DNA序列克隆到检索载体中以形成DNA载体复合物; (d)使DNA-载体复合物线性化; 和(e)使用同源重组将来自DNA-载体复合物的第一个DNA片段插入第二个DNA片段以形成组合的DNA分子; 和(f)除去选择性标记,从而产生组合的DNA分子。 本发明还提供了用于检索并将选定的DNA分子插入靶DNA分子的载体。

    Method for constructing and modifying large DNA molecules
    2.
    发明申请
    Method for constructing and modifying large DNA molecules 有权
    构建和修饰大DNA分子的方法

    公开(公告)号:US20050084970A1

    公开(公告)日:2005-04-21

    申请号:US10892579

    申请日:2004-07-16

    CPC分类号: C12N15/10

    摘要: This invention provides a method for combining overlapping DNA molecules comprising: (a) providing first and second DNA fragments, the first having a region homologous to a region in the second; (b) tagging the first DNA fragment with a selectable marker; (c) cloning the first DNA sequence into a retrieval vector to form a DNA-vector complex; (d) linearizing the DNA-vector complex; and (e) inserting the first DNA fragment from the DNA-vector complex into the second DNA fragment using homologous recombination to form a combined DNA molecule; and (f) removing the selectable marker, thereby generating a combined DNA molecule. The invention further provides a vector for retrieving and inserting a selected DNA molecule into a target DNA molecule.

    摘要翻译: 本发明提供了一种组合重叠DNA分子的方法,包括:(a)提供第一和第二DNA片段,第一DNA片段具有与第二DNA片段中的区域同源的区域; (b)用选择标记标记第一个DNA片段; (c)将第一DNA序列克隆到检索载体中以形成DNA载体复合物; (d)使DNA-载体复合物线性化; 和(e)使用同源重组将来自DNA-载体复合物的第一个DNA片段插入第二个DNA片段以形成组合的DNA分子; 和(f)除去选择性标记,从而产生组合的DNA分子。 本发明还提供了用于检索并将选定的DNA分子插入靶DNA分子的载体。