中文摘要：

根据小鼠肌肉生长抑制素（MSTN）mRNA序列设计4对shRNA序列,通过退火连接到shRNA过表达载体pGPU6/GFP/Neo,构建相应的过表达载体pGPU6/GFP/Neo-MSTN shRNA。转染C2C12细胞后,利用实时荧光定量PCR（QRT-PCR）及Western blot试验对shRNA基因沉默后MSTN mRNA及蛋白的表达水平进行检测。结果表明：设计并构建的4个MSTN shRNA过表达载体,在细胞水平都能有效降低MSTN mRNA及蛋白的表达水平,其中转染pGPU6/GFP/Neo-MSTN714载体细胞组基因沉默效率最高,MSTN mRNA和蛋白表达分别下降72%、74%,差异极显著（P〈0.01）。该研究成功设计了4个MSTN shRNA序列并构建了相应的高效表达载体,为后续通过调控MSTN表达研究其在肌肉发育中功能奠定基础。

英文摘要：

In this study,4pairs of MSTN shRNA were designed based on MSTN mRNA sequence and 4MSTN shRNA overexpression vectors pGPU6/GFP/Neo-MSTN shRNAs were constructed.After the vectors transfection to C2C12 cells,the MSTN mRNA and protein were detected with QRT-PCR and western blot methods.The results showed that4 MSTN shRNA overexpression vectors were constructed successfully;they all reduced the MSTN expression.The cells transfected with pGPU6/GFP/Neo-MSTN714 showed a higher gene silencing efficiency,MSTN mRNA and protein expression level was reduced by 72% and 74% respectively,the difference was significant（P〈 0.01）.Our study constructed the MSTN shRNAs and the overexpression vectors and established a foundation for studying the function of MSTN on muscle development.