基于Cre/loxP系统的糖皮质激素受体基因条件性打靶嵌合鼠的获得

目的:应用Cre/loxP系统建立诱导型糖皮质激素受体(GR)基因剔除小鼠模型,为在体内直接进行GR基因的研究提供条件.方法:针对GR基因第2外显子构建诱导型目标载体,将打靶载体电穿孔转染胚胎干细胞(embryonic stem cell,ES),经Southern杂交筛选发生正确同源重组的ES细胞,囊胚注射法制备嵌合体并进行生殖系检测及纯化建系.结果:成功构建了针对GR基因第2外显子的诱导型目标载体,转染ES细胞获得了工程化的ES细胞,经囊胚注射获得了由工程化的ES细胞和体细胞共同发育而来的嵌合体小鼠.结论:基于Cre/loxP系统的GR基因条件性打靶嵌合鼠的获得,为得到理想的诱导型GR基因剔除小鼠模型奠定了基础.

Generation of chimeras of glucocorticoid receptor conditional gene targeting mice based on Cre/loxP system

Objective:To establish an inducible model of glucocorticoid receptor (GR) gene knock-out mice using Cre/loxP system. Methods: An inducible targeting vector aiming at the exon 2 of GR gene was constructed. The targeting vector was electroporated into ES cells and targeted ES clones were obtained by Southern blot screening. Chimeras were acquired by blastocysts injection and the germ-line chimeras were obtained with the detection of gonad. Results: An inducible targeting vector aiming at the exon 2 of GR gene was successfully constructed. GR targeted ES cells were obtained by Southern blot screening and chimeras were produced by blastocysts injection. Conclusion: Generation of chimeras of GR conditional gene targeting mice based on Cre/loxP system has laid a solid foundation for establishing an ideal inducible model of GR gene konck-out mice.