水泡性口炎病毒核蛋白基因在大肠杆菌中的表达和鉴定

Expression of nucleoprotein gene of vesicular stomatitis virus indiana type in Escherichia coli

根据已经发表的IN型水泡性口炎病毒(VSV)核蛋白(N)基因的序列设计1对特异引物,应用RT-PCR技术扩增编码IN VSV的N基因,将PCR产物按相应的阅读框架克隆到表达载体pET-32a。并将重组质粒转化入大肠杆菌 BL21株,以1．0 mmol·L-1 IPTG在37℃的条件下诱导,N基因得到了高效表达。经过聚丙烯酰胺凝胶电泳以及用 VSV阳性血清进行Western blotting试验,结果表明所表达的融合蛋白产物分子质量与预期的65．3 ku相符。

The Nucleoprotein gene of vesicular stomatitis virus Indiana type was amplified by reverse transcription polymerase chain reaction （RT-PCR）. The DNA fragment was ligated with plasmid pET-32a and transformed into Eseheriehia Coli. The constructed recombinant plasmid was confirmed by RE digestion and sequencing, and induced with IPTG. The expressed protein was valued by SDS-PAGE and Western blot, and the results showed molecular weight of the recombinant protein in accordance with the expected 65.3 ku.