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.
出处
《广东农业科学》
CAS
CSCD
北大核心
2013年第17期133-136,共4页
Guangdong Agricultural Sciences
基金
国家转基因生物新品种培育科技重大专项(2011ZX08009-004)
贵州省科技厅农业攻关项目(黔科NY字[2012]3008号)
贵州大学研究生创新基金(校农科2012007)
