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由表达Cre重组酶的腺病毒载体介导的人类染色体中loxP位点间的高频重组。

High frequency recombination between loxP sites in human chromosomes mediated by an adenovirus vector expressing Cre recombinase.

作者信息

Wang P, Anton M, Graham F L, Bacchetti S

机构信息

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

出版信息

Somat Cell Mol Genet. 1995 Nov;21(6):429-41. doi: 10.1007/BF02310209.

Abstract

An adenovirus vector (AdCre1) expressing Cre recombinase has been used to induce recombination between loxP sites in human chromosomes. G418 resistant cells with one loxP site, generated by transfection with a plasmid containing loXp between the SV40 promoter and the G418 resistance (neo) gene, were infected with AdCre1 and transfected with a plasmid containing loxP adjacent to a promoterless hisD gene. This resulted in integration of hisD downstream of the SV40 promoter with gain of histidinol and loss of G418 resistance. Since AdCre1 is non-replicating and Cre expression transient, histidinol resistant cells containing the hisD gene flanked by loxP sites were stable. Reinfection of these cells with AdCre1 induced excision of hisD in over 90% of infected cells. This high efficiency of site-specific recombination suggests that AdCre1 may be exploited for temporal and tissue-specific regulation of gene expression and for chromosome engineering in vitro and in animals.

摘要

一种表达Cre重组酶的腺病毒载体(AdCre1)已被用于诱导人类染色体上loxP位点之间的重组。通过用一个在SV40启动子和G418抗性(neo)基因之间含有loXp的质粒转染产生的带有一个loxP位点的G418抗性细胞,用AdCre1进行感染,并转染一个在无启动子的hisD基因附近含有loxP的质粒。这导致hisD整合到SV40启动子的下游,同时获得组氨醇抗性并丧失G418抗性。由于AdCre1是非复制型的且Cre表达是瞬时的,含有被loxP位点侧翼的hisD基因的组氨醇抗性细胞是稳定的。用AdCre1再次感染这些细胞可在超过90%的感染细胞中诱导hisD的切除。这种高效的位点特异性重组表明,AdCre1可用于基因表达的时间和组织特异性调控以及体外和动物体内的染色体工程。

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