重组人乳头瘤病毒CTL表位疫苗的构建与表达

目的：构建人乳头瘤病毒的细胞毒性T淋巴细胞（CTL）表位疫苗并在大肠杆菌中表达。方法：应用多轮PCR 的方法合成CTL表位基因序列,将其亚克隆入pET28a表达载体中,并在大肠杆菌BL21 (DE3) 中表达重组蛋白质,用Western blotting对重组蛋白进行鉴定。结果：采用计算机表位预测的方法选取HPV6、11、16型的CTL表位，通过5轮PCR 人工合成人乳头瘤病毒CTL表位疫苗的基因片段HPVepi，用亚克隆的方法构建了重组表达质粒pET28a-Tat-HPVepi，并在大肠杆菌高效表达了该重组蛋白质，经Western blotting鉴定显示出特异性条带。结论：结合计算机表位预测和PCR的方法构建并在大肠杆菌中高效表达了一种人乳头瘤病毒CTL表位疫苗。

Construction and expression of human papillomavirus CTL epitope vaccine

Objective To construct a CTL epitope vaccine against human papillomavirus and to express it in E.coli.Methods The gene sequences encoding CTL epitopes selected by computer-assisted epitope prediction were synthesized by PCR and sub-cloned into pET28a expression vector.The recombinant plasmid pET28a-Tat-HPVepi was transformed to E.coli for expression and the expressed protein was identified by Western blotting.Results The DNA sequences of selected CTL epitopes were obtained by 5 rounds of PCR,the constructed recombinant plasmid pET28aTat-HPVepi was effectively expressed in E.coli.The expressed products,identified by Western blotting,showed specific bands.Conclusion The CTL epitope vaccine against human papillomavirus is successfully constructed and express in E.coli.