SARS-Co V S基因重组质粒的构建与瞬时表达

Construction and Transient Expression of SARS-Co V S Gene Recombinant Plasmid

根据GenBank中登录的SARS-COV(BJ01株)基因序列设计引物,采用反转录聚合酶链反应(RT-PCR)方法从SARS病毒中扩增得到约3 768 bp的片段,将其克隆到真核表达质粒pVAX1,鉴定正确后,命名为pVAX1/S。体外转染Hela细胞,间接免疫荧光鉴定S蛋白的表达。构建的重组质粒经酶切及测序鉴定,开放阅读框正确;间接免疫荧光方法鉴定该重组质粒能在Hela细胞中表达。

According to the data in Genebank, the specific primers of the SARS-CoV S gene was designed. The S gene was amplified from SARS-CoV by R＇F-PCR. Then it was inserted into the eukaryotic expression vector pVAX1, named pVAX1/S. After transfecting Hela cell with pVAX1/S, the expression of S protein was detected with indirect immunofluorescent method. The result indicated that recombinant plasmid of pVAX1/S was constructed successfully. Expression of S protein was detected by indirect immunofluorescent method.