HPV16E6真核表达载体的构建及其在CaSki细胞中的表达及定位  

Construction of HPV16E6 eukaryotic expression vector and its expression and location in CaSki cells

作　　者：王青元[1] 周颖[1] 杨霞[1] 李兵[1] 陶峰[1] 朱圆圆[1] 凌斌[1] 

机构地区：[1]安徽医科大学附属省立医院妇产科,安徽合肥230001

出　　处：《中国妇幼保健》2011年第31期4904-4906,共3页Maternal and Child Health Care of China

基　　金：国家自然科学基金〔8107212781001168〕;安徽高校省级自然科学研究项目〔KJ2010B375〕;安徽省自然科学基金面上项目〔090413117〕

摘　　要：目的:构建pEGFP-C2-HPV16E6真核表达载体,转染CaSki细胞,检测其在CaSki细胞中的表达。方法:用RT-PCR法从CaSki细胞中扩增出带有HindⅢ、XbaⅠ酶切位点的HPV16E6基因片段,将HPV16E6基因全长片段克隆到增强型绿色荧光蛋白(EGFP)基因真核表达载体pEGFP-C2上,通过脂质体将pEGFP-C2-HPV16E6转染入CaSki细胞。结果:pEGFP-C2-HPV16E6真核表达载体构建成功,转染CaSki细胞48h后经RT-PCR及Western-blot检测可见HPV16E6蛋白的高表达,激光共聚焦显微镜观察融合蛋白主要在CaSki细胞核内表达。结论:成功构建了pEGFP-C2-HPV16E6真核表达载体,为进一步对HPV16E6的研究奠定了基础。Objective：To construct pEGFP-C2-HPV16E6 eukaryotic expression vector,transfect CaSki cells,and detect its expression in CaSki cells.Methods：RT-PCR was used to amplify HPV16E6 gene segment in CaSki cells with HindⅢ and XbaⅠ restriction enzyme cutting sites,the total HPV16E6 gene segment was cloned into eukaryotic expression vector pEGFP-C2 encoding enhanced green fluorescent protein（EGFP）.HPV16E6 was transfected into CaSki cells according to plasmid.Results：pEGFP-C2-HPV16E6 eukaryotic expression vector was successfully constructed.High expression of 16E6 protein could be detected by RT-PCR and Western blot after transfecting into CaSki cells for 48 hours,the expression of fusion protein in CaSki cell nucleus was observed under laser scanning confocal microscope.Conclusion：pEGFP-C2-HPV16E6 eukaryotic expression vector is successfully constructed,which provide a foundation for further investigation of HPV16E6.

关 键 词：CASKI细胞 HPV16E6 真核表达载体 

分 类 号：R382.31[医药卫生—医学寄生虫学]