HPV 6b L1基因原核表达系统的构建及鉴定

克隆、构建人乳头瘤病毒6b型(HPV6b)L1基因重组质粒,并进行表达和鉴定。用PCR方法,从尖锐湿疣标本中扩增出HPV6b型L1基因,构建重组克隆质粒pblue-HPV6bL1,测序分析其基因变异。将本地株HPV6b型L1基因酶切后连接到原核表达质粒pBAD,构建原核表达系统pBAD-HPV6bL1/Top10,酶切鉴定证明重组质粒的正确性。经L-Arobinose诱导后表达HPV6b型L1蛋白,利用SDS-PAGE对表达产物进行鉴定。经酶切及序列分析鉴定,重组质粒pBAD-HPV6bL1构建成功,诱导后能够表达L1融合蛋白。HPV6bL1重组质粒构建成功,并获得HPV6bL1蛋白,为该蛋白的功能及HPV的基因工程疫苗研究提供了物质基础。

Construction and Identification of Prokaryotic Expression System with L1 Gene of Human Papillomavirus HPV 6b

L1 gene of human papillomavirus type 6b (HPV6b) was cloned into pBLUE plasmid to construct a recombinant one, and identified its expression. The HPV6b L1 gene was amplified from sample of condyloma accuminatum (CA) by PCR. L1 gene was also cloned into vector pBAD to form pBAD-HPV6bL1 plasmid and transferred into E.coli TOP10. The L1 protein was expressed after induced by L-arabinose and detected with SDS-PAGE. The recombinant plasmid was confirmed by restriction endonucleases digestion and sequencing. Therefore, L1 protein of HPV6b could be expressed in prokaryotic system. Thus provide material foundation to study the function of the protein and HPV vaccine engineering of CA.