-
公开(公告)号:US07781190B2
公开(公告)日:2010-08-24
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。
-
公开(公告)号:US20050084970A1
公开(公告)日:2005-04-21
申请号:US10892579
申请日:2004-07-16
申请人: Jian-Dong Huang , Xin-Mei Zhang , Julian Tanner
发明人: Jian-Dong Huang , Xin-Mei Zhang , Julian Tanner
IPC分类号: C12N15/10 , C12N15/867 , C12P19/34
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。
-