Sakai K, Mitani K, Miyazaki J
Department of Disease-related Gene Regulation Research (Sandoz), University of Tokyo, Japan.
Biochem Biophys Res Commun. 1995 Dec 14;217(2):393-401. doi: 10.1006/bbrc.1995.2789.
We have constructed an E1-defective adenovirus (Ad) vector designated AdCAG-Cre containing the Cre recombinase gene derived from bacteriophage P1 under control of the cytomegalovirus immediate early enhancer-chicken beta-actin hybrid (CAG) promoter. We examined the Cre-loxP-based recombination by this Ad vector in C2C12 cells bearing a reporter gene construct CAG-CAT-Z, which directs expression of the E. coli lacZ gene upon Cre-mediated excision of the CAT gene located between the CAG promoter and the lacZ gene. Nearly 100% of these cells were shown to start to produce beta-galactosidase after infection with the AdCAG-Cre vector at MOI 100. On the basis of this result, we discuss the possible use of the AdCAG-Cre vector to manipulate the gene expression in mammalian cells.
我们构建了一种E1缺陷型腺病毒(Ad)载体,命名为AdCAG-Cre,其包含源自噬菌体P1的Cre重组酶基因,该基因受巨细胞病毒立即早期增强子-鸡β-肌动蛋白杂交(CAG)启动子的控制。我们在携带报告基因构建体CAG-CAT-Z的C2C12细胞中检测了这种Ad载体基于Cre-loxP的重组,该构建体在Cre介导切除位于CAG启动子和lacZ基因之间的CAT基因后指导大肠杆菌lacZ基因的表达。在感染复数(MOI)为100的情况下,用AdCAG-Cre载体感染后,近100%的这些细胞开始产生β-半乳糖苷酶。基于这一结果,我们讨论了AdCAG-Cre载体在操纵哺乳动物细胞基因表达方面的可能用途。