轮状病毒VP7基因的克隆与表达

Cloning and Expression of Protein VP7 of Rotavirus

目的构建轮状病毒VP7的重组表达质粒,在大肠杆菌中对其进行表达。方法克隆编码轮状病毒VP7的基因,并在原核表达系统中表达了带有6×His标签的融合蛋白。结果经双酶切鉴定和基因测序证明成功重组了轮状病毒VP7基因的pQE-30重组质粒;蛋白质PAGE电泳表明,获得了分子量约为48000Mr的蛋白质。结论本研究获得了重组的轮状病毒VP7蛋白。

Objective To clone and express the VP7 gene of rotavirus in E.coli. Method The VP7 fragment was amplified by PCR and cloned into the prokaryotic expression vector pQE-30 vector. After verified by digesting with restriction endonuclease and sequencing. The recombinant vector was transformed into E.coli BL21. The expressed protein was detected by SDS-PAGE. Result According to the SDS-PAGE analysis, the relative molecular mass（Mr） of the fusion protein is about 48000 Mr. Conclusion The VP7 of rotavirus was cloned and expressed as fusion protein in E.coli.