HPV16E6羧基端编码基因的克隆及真核细胞表达

GENE CLONING AND EUKARYOTIC EXPRESSION OF HPV16E6 C-TERMINAL REGION

目的 :探讨用人乳头瘤病毒 16型 (HPV16)早期基因E6羧基端基因片段 (E6C)研制DNA疫苗的可行性。方法 :采用PCR方法从质粒 pHPV16中获得E6C端基因片段 ,将其克隆入含巨细胞病毒(CMV)启动子的真核表达载体 ,脂质体介导基因转染LA795小鼠肺腺癌细胞并检测E6C的表达。结果 :成功构建了真核表达质粒 pLNCE6C ,免疫组化结果显示真核表达质粒pLNCE6C在LA795细胞中获得有效表达。结论 :选择去除转化功能区的E6C端基因构建真核表达质粒是一有益的尝试 。

Objectives:To explore the feasibility of utilizing the gene fragment of Cterminal region of human papillomavirus type 16 E6(HPV16 E6)for DNA vaccine.Methods:E6C gene fragment was amplified from genome of pHPV16 by PCR,and cloned into eukaryotic plasmid pLNCX,then teansfected LA795 cell with the recombinant plasmid pLNCE6C using lipofectamine reagent.Results:The result showed recombinant pLNCE6C was successfully constructed and the E6 protein was detected by immunohistochemistry in the transfected cells.Conclusion:This study provides a good basis for further research on HPV16 E6C DNA vaccine.