Zeh K, Andahazy M, O'Gorman S, Baribault H
The Burnham Institute, 10901 N. Torrey Pines Road, La Jolla, CA 92037, USA and Gene Expression Laboratory,The Salk Institute, 10010 N. Torrey Pines Road, La Jolla, CA 92037, USA.
Nucleic Acids Res. 1998 Sep 15;26(18):4301-3. doi: 10.1093/nar/26.18.4301.
Deletion of genes in defined cell types has been achieved using a combination of gene targeting techniques and the Cre- lox P recombination system. Here we present a method to selectively isolate genetically altered primary cell cultures based on the permanent activation of a drug-resistance gene by the Cre recombinase. Transgenic mice were generated harboring a dormant form of the hygromycin resistance gene. This mouse line was crossed with mice carrying a constitutive Cre gene and an endogenous floxed allele. Primary fibroblasts established from triple transgenic embryos displayed not only hygromycin resistance but also recombination of the endogenous floxed allele. These results prove the potential of this approach.
利用基因靶向技术和Cre-loxP重组系统的组合,已实现了在特定细胞类型中删除基因。在此,我们提出一种基于Cre重组酶对耐药基因的永久激活来选择性分离基因改变的原代细胞培养物的方法。构建了携带潮霉素抗性基因休眠形式的转基因小鼠。将该小鼠品系与携带组成型Cre基因和内源性loxP侧翼等位基因的小鼠杂交。从三联转基因胚胎建立的原代成纤维细胞不仅表现出潮霉素抗性,还表现出内源性loxP侧翼等位基因的重组。这些结果证明了该方法的潜力。