| 刘敏，许厚强，陈伟，陈祥，李飞.关岭黄牛MSTN启动子真核报告载体的构建与启动活性研究[J].广东农业科学,2013,40(17):133-136 |
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关岭黄牛MSTN启动子真核报告载体的构建与启动活性研究 |
| Construction and promotion research of MSTN reporter plasmid in Guanling cattle |
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| DOI： |
| 中文关键词: MSTN 基因 启动子 荧光素酶 C2C12 3T3-L1 |
| 英文关键词: MSTN gene promoter luciferase C2C12 3T3-L1 |
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| 摘要点击次数: 1589 |
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| 中文摘要: |
| 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-Basic- MSTN-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遥测序结果表明，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 倍尧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。 |
| 英文摘要: |
| 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-promoter 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. |
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