Kanegae Y, Lee G, Sato Y, Tanaka M, Nakai M, Sakaki T, Sugano S, Saito I
Laboratory of Molecular Genetics, University of Tokyo, Japan.
Nucleic Acids Res. 1995 Oct 11;23(19):3816-21. doi: 10.1093/nar/23.19.3816.
A recombinant adenovirus (Ad) expressing Cre recombinase derived from bacteriophage P1 was constructed. To assay the Cre activity in mammalian cells, another recombinant Ad bearing an on/off-switching reporter unit, where a LacZ-expression unit can be activated by the Cre-mediated excisional deletion of an interposed stuffer DNA, was also constructed. Co-infection experiments together with the Cre-expressing and the reporter recombinant Ads showed that the Cre-mediated switching of gene expression was detected in nearly 100% of cultured CV1, HeLa and Jurkat cells. These results suggest that the recombinant Ad efficiently expressed functional Cre and offers a basis for establishing a powerful on/off switching strategy of gene expression in cultured mammalian cells and presumably in transgenic animals. The method is also applicable to construction of recombinant Ad bearing a gene the expression of which is deleterious to propagation of recombinant Ad.
构建了一种表达源自噬菌体P1的Cre重组酶的重组腺病毒（Ad）。为了检测哺乳动物细胞中的Cre活性，还构建了另一种携带开/关切换报告单元的重组Ad，其中LacZ表达单元可通过Cre介导的插入填充DNA的切除缺失而被激活。将表达Cre的重组Ad与报告重组Ad一起进行共感染实验表明，在几乎100%的培养CV1、HeLa和Jurkat细胞中检测到了Cre介导的基因表达切换。这些结果表明，重组Ad能高效表达功能性Cre，并为在培养的哺乳动物细胞以及可能在转基因动物中建立强大的基因表达开/关切换策略提供了基础。该方法也适用于构建携带对重组Ad增殖有害的基因的重组Ad。
