公开(公告)号：US20040166528A1

公开(公告)日：2004-08-26

申请号：US10817950

申请日：2004-04-06

申请人： Biogen Idec Inc.

发明人： Mitchell R. Reff ,  Richard Spence Barnett ,  Karen Retta McLachlan

IPC分类号： C12Q001/68 ,  C12N015/85

CPC分类号： C07K16/2851 ,  C07K14/70521 ,  C07K16/2896 ,  C07K2319/02 ,  C12N15/85 ,  C12N15/907 ,  C12N2800/108 ,  C12N2830/42 ,  C12N2840/20 ,  C12N2840/44

摘要： A method for achieving site specific integration of a desired DNA at a target site in a mammalian cell via homologous recombination is described. This method provides for the reproducible selection of cell lines wherein a desired DNA is integrated at a predetermined transcriptionally active site previously marked with a marker plasmid. The method is particularly suitable for the production of mammalian cell lines which secrete mammalian proteins at high levels, in particular immunoglobulins. Novel vectors and vector combinations for use in the subject cloning method are also provided.

摘要翻译： 描述了通过同源重组在哺乳动物细胞的靶位点实现所需DNA的位点特异性整合的方法。 该方法提供细胞系的可重复选择，其中所需DNA整合在预先标记有标记质粒的转录活性位点上。 该方法特别适用于生产以高水平分泌哺乳动物蛋白质的哺乳动物细胞系，特别是免疫球蛋白。 还提供了用于本发明克隆方法的新型载体和载体组合。