GFP-HPV16 E5融合蛋白在HeLa细胞内的表达与定位  

Expression and Localization of GFP-HPV16 E5 Fusion Protein in HeLa Cells

作　　者：唐双阳[1] 陈杰[1] 李薇[1] 周良[1] 余敏君[1] 朱翠明[1] 万艳平[1] 

机构地区：[1]南华大学病原生物学研究所,湖南衡阳421001

出　　处：《南华大学学报（医学版）》2010年第6期733-735,共3页Journal of Nanhua University(Medical Edition)

基　　金：国家自然科学基金项目(30973402).

摘　　要：目的研究GFP-HPV16 E5融合蛋白在HeLa细胞内的表达与定位。方法 PCR扩增HPV16E5基因片段,将其连接到真核表达载体pEGFP-C1,双酶切及测序鉴定,构建表达载体pEGFP-C1/HPV16 E5;将质粒pEGFP-C1/HPV16 E5和pEGFP-C1分别转染HeLa细胞,在倒置荧光显微镜下观察GFP-HPV16 E5融合蛋白在HeLa细胞内的表达与定位。结果 GFP荧光均匀分布在细胞浆和细胞核;GFP-HPV16 E5融合蛋白组荧光分布在细胞浆。结论 GFP-HPV16 E5融合蛋白能在HeLa细胞内表达且定位于细胞浆。Objective To investigate the expression and localization of GFP - HPV16 E5 fusion proteins in HeLa cells.Methods HPV16 E5 gene fragment amplified by PCR was inserted into eukaryotic expression vector of pEGFP -C1 with the same restriction sites.Enzymes digestion and DNA sequencing were used to identify E5 gene fragment.The eukaryotic expression vector of pEGFP - C1/HPV16 E5 or pEGFP - C1 was transfected into HeLa cells,respectively.The expression and localization of GFP - HPV16 E5 or GFP was observed under fluorescent microscope.Results Fluorescence could be observed not only in the cytoplasma but also in the nuclei of the cells transfected by pEGFP - C1,and only in the cytoplasma of the cells transfected by pEGFP - C1/HPV16 E5.Conclusion Eukaryotic expression vector of pEGFP - C1/HPV16 E5 successfully expressed GFP - HPV16 E5 fusion proteins in HeLa cells and located in the cytoplasma.

关 键 词：人乳头瘤病毒16型 E5 定位 

分 类 号：R373[医药卫生—病原生物学]