犬新孢子虫NcSRS2基因片段的克隆及原核表达

Cloning and Prokaryotic Expression of NcSRS2 Gene of Neospora caninum

以含有犬新孢子虫NcSRS2基因的质粒pMD18-NcSRS2为模板,应用PCR方法扩增Nc-SRS2基因,将该基因片段克隆至原核表达载体pGEX-4T-2,构建了重组表达质粒pGEX-Nc-SRS2,转化大肠杆菌BL21中并诱导表达。结果显示,克隆的基因片段长1100bp,SDS-PAGE、Western blotting分析显示,重组质粒pGEX-NcSRS2在大肠杆菌中得到了高效表达。

The NcSRS2 gene fragment of Neospora caninum was amplified from pMD18-NcSRS2 by PCR,and then inserte into the pGEX--4T--2 vector. The recombinant expression plasmid pGEX--NcSRS2 was successfully constructed. The gene was induced by IPTG to express in Escherichia coli BL21. Results showed that the NcSRS2 cDNA was 1 100bp in length, SDS-PAGE.Western blotting analysis revealed that recombinant pGEX-NcSRS2 has been highly expressed in E. coli.