公开(公告)号：US06379943B1

公开(公告)日：2002-04-30

申请号：US09263650

申请日：1999-03-05

申请人： Frank L. Graham ,  Robin J. Parks ,  Philip Ng

发明人： Frank L. Graham ,  Robin J. Parks ,  Philip Ng

IPC分类号： C12N510

CPC分类号： C12N7/00 ,  C12N15/86 ,  C12N15/907 ,  C12N2710/10343 ,  C12N2710/10351 ,  C12N2800/30

摘要： In the present invention, viruses, plasmids or both are constructed which contain viral DNA and lox sites positioned such that site-specific recombination between lox sites in separate plasmids results in generation of infectious viral DNA at high-efficiency in cotransfected host cells that have been engineered to express the Cre recombinase. Because of the high-efficiency and specificity of the Cre enzyme, suitably engineered plasmids can be readily recombined to produce infectious virus at high-efficiency in cotransfected 293 cells, without, at the same time, producing wild-type adenovirus, with the attendant problems for removal thereof. Use of recombinases besides Cre and recombinase recognition sites besides lox sites, and use of cells other than 293 cells are also disclosed and enabled, as are kits incorporating the site-specific vector system, as well as compositions and methods for using such compositions as vaccines or in gene therapeutic applications.

摘要翻译： 在本发明中，构建了含有病毒DNA和lox位点的病毒，质粒或两者，其位置使得分离的质粒中的lox位点之间的位点特异性重组导致在共转染的宿主细胞中高效率地产生感染性病毒DNA 旨在表达Cre重组酶。 由于Cre酶的高效率和特异性，合适的工程改造质粒可以在共转染293细胞中高效率地重组，产生感染性病毒，同时产生野生型腺病毒，伴随着问题 用于去除。 除了lox位点之外的Cre和重组酶识别位点以及使用除293细胞之外的细胞的重组酶的使用也被公开和启用，以及包含位点特异性载体系统的试剂盒以及使用这些组合物作为疫苗的组合物和方法 或在基因治疗应用中。

公开(公告)号：US07132290B2

公开(公告)日：2006-11-07

申请号：US09909414

申请日：2001-07-19

申请人： Frank L. Graham ,  Robin J. Parks ,  Philip Ng

发明人： Frank L. Graham ,  Robin J. Parks ,  Philip Ng

IPC分类号： C12N15/87 ,  C12N15/861

CPC分类号： C12N7/00 ,  C12N15/86 ,  C12N15/907 ,  C12N2710/10343 ,  C12N2710/10351 ,  C12N2800/30

摘要： In the present invention, viruses, plasmids or both are constructed which contain viral DNA and lox sites positioned such that site-specific recombination between lox sites in separate plasmids results in generation of infectious viral DNA at high-efficiency in cotransfected host cells that have been engineered to express the Cre recombinase. Because of the high-efficiency and specificity of the Cre enzyme, suitably engineered plasmids can be readily recombined to produce infectious virus at high-efficiency in cotransfected 293 cells, without, at the same time, producing wild-type adenovirus, with the attendant problems for removal thereof. Use of recombinases besides Cre and recombinase recognition sites besides lox sites, and use of cells other than 293 cells are also disclosed and enabled, as are kits incorporating the site-specific vector system.

摘要翻译： 在本发明中，构建了含有病毒DNA和lox位点的病毒，质粒或两者，其位置使得分离的质粒中的lox位点之间的位点特异性重组导致在共转染的宿主细胞中高效率地产生感染性病毒DNA 旨在表达Cre重组酶。 由于Cre酶的高效率和特异性，合适的工程改造质粒可以在共转染293细胞中高效率地重组，产生感染性病毒，同时产生野生型腺病毒，伴随着问题 用于去除。 除了含有lox位点之外的Cre和重组酶识别位点以及使用除293细胞之外的细胞的重组酶的使用也被公开和启用，以及包含位点特异性载体系统的试剂盒。