作　　者： ; ; ; ; ;

机构地区： 贵州大学生命科学学院

出　　处： 《广东农业科学》 2013年第17期133-136,共4页

摘　　要： 采用PCR技术扩增牛MSTN基因启子,亚克隆至荧光素酶表达载体pGL3-Basic中,构建重组报告载体pGL3-BasicMSTN-promoter。将重组报告载体pGL3-Basic-MSTN-promoter与内参质粒pRL-TK用脂质体法瞬时共转染小鼠成肌细胞系C2C12和小鼠胚胎成纤维细胞3T3-L1,通过双荧光素酶活性检测其启动子活性。测序结果表明,成功构建了牛MSTN基因真核报告载体pGL3-Basic-MSTN-promoter,瞬时转染试验表明,pGL3-Basic-MSTN-promoter在C2C12细胞和3T3-L1细胞中的启动子活性分别为pGL3-Basic空载体的14.53倍、5.02倍。研究结果为进一步研究MSTN基因的表达调控机制奠定了基础。 The MSTN promoter was amplified and inserted into luciferase pGL3-Basic vector to construct recombined pGL3-Basic- MSTN-promoter reporter plasmid. Transient transfection was performed in mouse myoblasts cell line C2C12 and mouse embryonal fibroblast cell line 3T3-L1, and pRL-TK was used to determine the transfection efficiency. The sequencing results confirmed that the pGL3-Basic-MSTN-promotcr sequence was correct and expression in C2C12 and 3T3-L1 were 14.53 times and 5.02 tines more than pGL3-Basic plasmid respectively. The results could lay an experimental foundation for further studying the mechanism of MSTN expression and regulation.

关 键 词： 基因 启动子 荧光素酶

领　　域： [生物学]