人乳头瘤病毒16L1蛋白的原核表达及免疫原性研究

采用聚合酶链反应技术从宫颈癌组织中获得人乳头瘤病毒(HPV)16L1基因,克隆到pGEM—TEasy载体中,再连接到原核表达载体pGEX-6p-1谷胱甘肽-S-转移酶基因的下游,重组质粒转化大肠杆菌BL21,经IPTG诱导、SDS—PAGE分析,用表达产物免疫小鼠,获得的血清进行间接免疫荧光检测。结果表明：获得的pGEX-6p-1-L1基因在大肠杆菌中能以谷胱甘肽-S-转移酶融合蛋白的形式得到高效表达,表达产物免疫小鼠,间接免疫荧光显示免疫血清能与peDNA—L1转染的B16细胞呈现特异反应,说明体外表达的HPV16L1蛋白保留了天然蛋白的抗原性,可作为HPV的诊断抗原、免疫学分析和疫苗研制的原料。

Expression of human papillomavirous type 16L1 protein in a prokaryotic expression system and immunity original research

The PCR product was cloned into the pGEM-T Easy vector, then was inserted into downstream of GST in the pGEX-6p-1 vector to construct a recombinant plasmid pGEX-6p-1-L1. After the transformation of E. coli BL21 host bacterium, pGEX-6p-1-L1 was induced by IPTG and identified by SDS-PAGE. Immunized mice with the expression product acquired the serum, then conduct indirect fluorescence assay. The result of the indirect fluorescence assay showed that the serum of mice immunized with the expression product responsing to the B16 cells was transfected by pcDNA-L1. These data suggested that the expressed HPV16L1 protein in vitro reserved part of immunogenicity of natural protein, and could be used as the HPV antigen of diagnosis, the material of immunology analysis and the vaccine study.