扬州大学学报（农业与生命科学版）2016，Vol.37Issue(4)：1-4,4.DOI:10.16872/j.cnki.1671-4652.2016.04.001
小鼠MSTN shRNA可控表达载体构建及基因沉默效率研究
Construction of controllable mice MSTN shRNA expression vector and the gene silencing efficiency study
摘要
Abstract
Myostatin (MSTN),previously referred to as growth differentiation factor 8,is a negative regulator of skeletal muscle growth.In this study,a pair of MSTN shRNA was designed based on the MSTN sequence,and an integrated tetracycline induced expression vector,pSingle-tTS-MSTN shRNA-GFP,was constructed to express MSTN shRNA.The recombinant vector was transfected to C2C12 cell lines and screened with G418.MSTN shRNA expression was induced with DOX.The GFP expression was observed under a fluorescent microscope and the MSTN mRNA and protein were detected with QRT-PCR and western blot methods.The results showed that,after DOX inducing,the GFP can be observed and the MSTN mRNA decreased 70％ in the cells groups transfected with pSingle-tTS-MSTN shRNA-GFP vector cell groups,the difference between them is extremely significant (P＜0.01).The MSTN protein in this group decreased 90％ or so,whereas the other 3 groups showed no obvious difference(P＞0.05).A controllable MSTN shRNA overexpression vector was constructed successfully and showed a well-controlled gene expression efficiency.This study lays the foundation for prepairing MSTN shRNA controllable expression transgenic mice and the gene function analysis on muscle development.关键词
肌肉生成抑制素/可控表达/shRNA/基因沉默Key words
myostatin/controllable expression/shRNA/gene silencing分类
生物科学引用本文复制引用
杨跃飞,祁钰钰,杨开典,齐传翔,鞠辉明..小鼠MSTN shRNA可控表达载体构建及基因沉默效率研究[J].扬州大学学报（农业与生命科学版）,2016,37(4):1-4,4.基金项目
国家自然科学基金资助项目(31101683、31272405) （31101683、31272405）
江苏高校品牌专业建设工程项目(PPZY2015B158) （PPZY2015B158）
江苏省动物重要疫病与人兽共患病防控协同创新中心项目(2016-10) （2016-10）
