人微小病毒B19VP2蛋白抗原在大肠杆菌中的表达

Expression of human parvovirus B19 VP2 protein antigen in E. Coli

目的:利用原核系统表达人微小病毒B19的重组蛋白抗原,为建立ELISA方法诊断B19病毒感染提供特异性抗原。方法:采用PCR方法扩增B19VP2基因,将之插入到T载体中进行测序,验证序列正确后再重组入原核表达载体pET-30a(+),并转化大肠杆菌BL21(DE3)菌株,进行诱导表达。用SDSPAGE分析表达产物。表达产物经亲合层析柱纯化得到VP2重组蛋白抗原。用B19VP2IgG对该蛋白进行了Western-Blot分析。结果:筛选出2株B19VP2蛋白抗原高表达菌株,经IPTG诱导后,表达VP2重组蛋白量占菌体总蛋白量的24.6%。结论:VP2重组蛋白具有良好的抗原活性。

Aim: To express recombinant protein of human parvovirus B19 using prokaryotic expression system in order to provide specific antigen for diagnosis of human parvovirus B19 infection by ELISA.Methods: B19 VP2 gene was amplified using PCR, and then was inserted into T-vector and sequenced. The gene was recombined with prokaryotic expression vector pET-30a(+) after the sequence was identified to be right. BL21(DE3) was transformed with pET-30a(+)-VP2 and induced to express the protein. The expression product was analyzed by SDS-PAGE and purified by NI-NTA column chromatography, then VP2 recombinant protein antigen was gained. The protein was analyzed by B19 VP2 IgG through Western-Blot. Results: Two strains that could highly express B19 VP2 protein antigen were screened out. After induced by IPTG,the expression of recombinant protein occupied 24.6% in total protein. Conclusion: The recombinant protein of VP2 has better antigen activity.