人乳头瘤病毒16型中国株L1基因的体外真核表达

目的 构建人类乳头瘤病毒16型(HPV16)中国株晚期基因L1的真核表达系统.方法 将HPV16中国株L1基因从pCR2.1/HPV16L1定向亚克隆至pTacer-CMV载体,构建重组质粒pTaeer-CMV/HPV16L1,将重组质粒用脂质体转染N1H3T3、HeLa细胞,用透射电镜、SDS-PAGE、蛋白印迹等方法观察HPV16 L1蛋白的表达.结果 NIH3T3、HeLa细胞经pTacer-CMV/HPV16L1转染后,SDS-PAGE、蛋白印迹等实验显示可表达HPV16 L1蛋白,电镜下可见病毒样颗粒(VLP).结论 pTacer-CMV/HPV16L1构建成功,可在体外表达HPV16中国株L1蛋白.

Eukaryotic expression of human papillomavirus type 16 L1 gene in vitro

Objective To construct a plasmid for eukaryotic expression of later gene L1 of human papillo-mavims type 16(HPV16)Chinese strain.Methods,The L1 gene of HPV16 Chinese strain was subcloned from pCR2.1/HPV16L1 to pTracer-CMV.HeLa and NIH3T3 cells were transfeeted With pTracer-CMV/HPV16L1 by liposome.The expression of L1 protein was observed by transmission electronic microscope,SDS polyacrylamide gel electrophoresis(SDS-PAGE),and Western blotting.Results SDS-PAGE and Western blotting showed that HPV16 LI protein could be detected in the HeLa and NIH3T3 cells transfected by pTrracer-CMV/HPV16L1.The serf-assembled virus-like particles(VLP)could be observed by electronic microscope.Condmion pTracer-CMV/HPV16L1 had been successfully constructed,and could express the L1 protein of HPV16 Chinese strain in vitro.