猪卵泡抑素基因特异表达载体的构建及鉴定

Pig follistatin gene-specific expression vector and identification

采用RT-PCR方法扩增猪的FST基因完整编码区,采用双酶切和连接方法构建可在肌肉细胞中特异表达的真核载体,并转染C2C12细胞系进行验证。结果表明,猪FST基因全长1 032 bp,与网上已提交序列相似度为99.13%,成功构建可在肌肉细胞中特异表达的真核载体pEGFP-C1-α-actin-FST,转染C2C12细胞后,通过实时定量PCR检测,表明FST基因过表达可抑制MSTN基因表达。

RT-PCR was used to amplify the entire coding region of the FST gene pigs, With double enzymes and the connection method to build can be specific in the muscle cells of eukaryotic expression vector, and transfected C2C12cells for verification. The results showed that pigs FST gene was 1 032 bp, with the online submitted was 99.13% sequence similarity, successfully constructed a specific expression in the muscle cells of the eukaryotic expression vector pEGFP- C1- α- actin- FST,transfected C2C12cells, by Real-time quantitative PCR, showed overexpression FST could inhibit MSTN gene expression.