Yang Y S, Hughes T E
Yale University School of Medicine, New Haven, CT 06520, USA.
Biotechniques. 2001 Nov;31(5):1036, 1038, 1040-1. doi: 10.2144/01315st03.
The Cre/lox system is a powerful genetic tool with which to manipulate the genome. Here, we describe the development of a simple reporter system for Cre recombinase, called the Cre Stoplight. In the absence of Cre, the red fluorescent protein is expressed; when Cre catalyzes a recombination event, the green fluorescent protein is produced. Testing this system in transiently transfected cells showed that it produced robust signals (90% of the cells converted from red to green) when equal amounts of the plasmids encoding Cre recombinase and the Cre Stoplight were used. A 1:100 ratio of enzyme to reporter plasmid produced similar results, and a 1:10000 ratio was necessary to significantly reduce the number of cells converting to green (1%).
Cre/lox系统是一种用于操纵基因组的强大遗传工具。在此,我们描述了一种针对Cre重组酶的简单报告系统的开发,称为Cre Stoplight。在没有Cre的情况下,红色荧光蛋白表达;当Cre催化重组事件时,产生绿色荧光蛋白。在瞬时转染细胞中测试该系统表明,当使用等量编码Cre重组酶和Cre Stoplight的质粒时,它产生了强烈的信号(90%的细胞从红色转变为绿色)。酶与报告质粒1:100的比例产生了类似的结果,而1:10000的比例对于显著减少转变为绿色的细胞数量(1%)是必要的。