发明授权
US5925544A Method of homologous recombination followed by in vivo selection of DNA
amplification
失效
同源重组方法，然后进行DNA扩增的体内选择
- 专利标题： Method of homologous recombination followed by in vivo selection of DNA amplification
- 专利标题（中）： 同源重组方法，然后进行DNA扩增的体内选择
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申请号： US954391申请日： 1997-10-20
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公开(公告)号： US5925544A公开(公告)日： 1999-07-20
- 发明人: Steen Troels J.o slashed.rgensen
- 申请人： Steen Troels J.o slashed.rgensen
- 申请人地址： DKX Bagsvaerd
- 专利权人： Novo Nordisk A/S
- 当前专利权人： Novo Nordisk A/S
- 当前专利权人地址： DKX Bagsvaerd
- 主分类号： C12N15/64
- IPC分类号： C12N15/64 ; C12N15/90 ; C12P21/00 ; C07H21/04 ; C12P19/34 ; C12Q1/68
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.
公开/授权文献
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