发明名称 Method of homologous recombination followed by in vivo selection of DNA amplification
摘要 A method of amplifying in vivo a DNA sequence B present in a genome of a parent cell, comprising (a) integrating in a genome of said cell a DNA construct comprising the structure C-M-A-D, in which both A and C denote a DNA sequence which is homologous with a genomic DNA fragment either flanking or overlapping the DNA sequence B, the sequence C being located in the opposite end the sequence B as compared to A, D denotes a DNA sequence which is homologous with a genomic DNA fragment located distal for C as compared to B, and M denotes a DNA sequence encoding a selection marker, (b) selecting for cells in which the DNA sequence M has been integrated in the genome, which cells comprise, in any orientation, the structure A-B-C-M-A-D and (c) propagating the cells selected in step (b) under increasing selection pressure to obtain a cell which has obtained an increased number of genomically integrated copies of the DNA sequences B and M.
申请公布号 US5925544(A) 申请公布日期 1999.07.20
申请号 US19970954391 申请日期 1997.10.20
申请人 NOVO NORDISK A/S 发明人 JOERGENSEN, STEEN TROELS
分类号 C12N15/64;C12N15/90;(IPC1-7):C12P21/00;C12P19/34;C12Q1/68;C07H21/04 主分类号 C12N15/64
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