新型载体蛋白P64K在大肠杆菌中的可溶性表达与纯化

The soluble expression in E.coli and purification of a new carrier protein,P64K

用常规PCR方法从脑膜炎球菌中克隆出P64K基因,构建重组质粒pET28a-P64K转化BL21穴DE3雪宿主菌,经IPTG诱导表达筛选P64K蛋白的高表达菌株。结果证实P64K蛋白为胞内可溶性表达,表达量约占胞内总蛋白的30%。重组P64K蛋白经Butyl疏水柱、G200分子筛柱和Q阴离子柱三步层析后纯度达到95%以上,为今后进一步的功能和应用研究打下了良好的基础。

P64K gene was amplified from Neisseria meningitids by using PCR and fused to the plasmid pET28a,then the recombinant plasmid was transformed into host cell BL21(DE3)and the high expressive clone was screened by using ex-pression test of IPTG induction.P64K was expressed as high level as a soluble form,accounting for approximately30%of the total cell protein.After three step chromatograhpy comprising of butyl hydrophobic,G200gel filtration and Q anion ex-change,the P64K protein was purified to appromixately95%.