O型口蹄疫病毒VP1基因和牛集落刺激因子基因的融合表达

Fusion expression of foot-and-mouth disease virus type O VP1 gene with bovine GM-CSF gene

将O型口蹄疫病毒VP1基因和牛集落刺激因子(GMCSF)成熟肽基因共同克隆到原核表达载体pGEX6P1中,转化RS21后经IPTG诱导,实现了重组融合蛋白BoGMCSF/VP1在大肠埃希氏菌RS21中的高效表达,表达产物经SDSPAGE和Westernblotting分析,重组的蛋白质分子质量约为66ku,与预期大小相符。表达蛋白占菌体总蛋白的30%,表达产物以包涵体形式存在;包涵体提取物经变性复性,用透析方法提纯,得到高纯度的表达蛋白。

The VP1 gene of foot-and-mouth disease virus type O and the bovine GM-CSF gene were cloned into the prokaryotic expression vector pGEX-6P-1 ,and transformed into RS21 cells. The transformant was induced by IPTG, and the recombinant BoGM-CSF/VP1 was expressed as a fusion protein with a GST tag . The fusion protein was about 66ku in size and accounted for 30% of total protein. The recombinant protein aggregated to form insoluble bodies in RS21. The inclusion bodies were purified under denaturing and refolded conditions, then the purified BoGM-CSF/VP1 protein were acquired.