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表达位点特异性重组酶Cre的人293细胞系的构建与鉴定

Production and characterization of human 293 cell lines expressing the site-specific recombinase Cre.

作者信息

Chen L, Anton M, Graham F L

机构信息

Department of Biology, McMaster University, Hamilton, Ontario, Canada.

出版信息

Somat Cell Mol Genet. 1996 Nov;22(6):477-88. doi: 10.1007/BF02369439.

Abstract

We have constructed 293 cell lines expressing the site-specific Cre recombinase from bacteriophage P1, that acts on a 34 bp target sequence called loxP. Stably transformed cells were obtained by transfection with a plasmid containing Cre and a selectable marker under the control of viral promoters. The resulting 293Cre cell lines could be used to induce expression from adenovirus vectors containing reporter genes under the control of a Cre responsive "molecular switch." High efficiency recombination was observed for Ad viral DNA containing loxP sites. The Cre expressing cell lines described here are likely to be useful for several purposes: For expression of toxic gene products from Cre inducible viral vectors, to induce recombination between loxP sites in transfected plasmids, and to induce deletions or rearrangements of genes defined by loxP sites in viral genomes.

摘要

我们构建了表达来自噬菌体P1的位点特异性Cre重组酶的293细胞系,该重组酶作用于一个名为loxP的34bp靶序列。通过用含有Cre和在病毒启动子控制下的选择标记的质粒转染获得稳定转化的细胞。所得的293Cre细胞系可用于诱导在Cre响应性“分子开关”控制下含有报告基因的腺病毒载体的表达。对于含有loxP位点的腺病毒DNA,观察到了高效重组。本文所述的表达Cre的细胞系可能在多个方面有用:用于从Cre诱导型病毒载体表达毒性基因产物,诱导转染质粒中loxP位点之间的重组,以及诱导病毒基因组中由loxP位点定义的基因的缺失或重排。

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