人乳头瘤病毒16L1-E7蛋白的原核表达及分析纯化研究

Prokaryotic Expression and Purification of Human Papillomavirus 16L1-E7 Protein Based on Epitope Analysis

目的采用基于抗原表位分析对人乳头瘤病毒16L1-E7蛋白的原核表达和纯化进行分析,为相关疫苗的研究提供借鉴。方法将pET-32a/16L1-E7重组质粒转化BL21大肠杆菌,IPTG诱导16L1-E7蛋白表达,并对表达蛋白纯化后进行抗原特异性鉴定。结果重组质粒扩增后检测条带单一,菌落PCR和酶切片段位置一致;经0.1 mmol·L^(-1)的IPTG诱导后重组质粒泳道目标蛋白表达明显增多,且抗原特异性好。结论成功诱导pET-32a/16L1-E7重组质粒表达并进行了纯化,获得了抗原特异性较好的目标蛋白。

Objective The prokaryotic expression and purification of human papillomavirus 16L1-E7 protein were analyzed based on antigen epitope analysis,in order to provide reference for the research of related vaccines.Methods The recombinant plasmid pET-32a/16L1-E7 was transformed into BL21 Escherichia coli and induced 16L1-E7 protein expression by IPTG.The expressed protein was purified for antigen specificity identification.Results After the recombinant plasmid was amplified,the detection bands were single,and the colony PCR and the location of the fragment was at the same location.After induction by 0.1 mmol·L-1 IPTG,the recombinant plasmid lane showed that the target protein expression increased significantly,and showed a good antigen-specificity.Conclusion The expression of pET-32a/16L1-E7 recombinant plasmid was successfully induced and purified,and the target protein with good antigen specificity was obtained.