HCV-F基因在hepG2细胞中的表达以及对PI3K的影响  

Expression of the HCV F Gene in HepG2 cells and the Effect on the Expression of PI3k

作　　者：储春丽[1] 邓小昭[2] 董莉莉[1] 胡学芳[2] 喻荣彬[1] 张云[2] 吴超[3] 

机构地区：[1]南京医科大学基础医学院,江苏南京210002 [2]南京军区疾病预防控制中心,江苏南京210002 [3]南京大学医学院附属鼓楼医院,江苏南京210029

出　　处：《药物生物技术》2008年第4期235-238,共4页Pharmaceutical Biotechnology

基　　金：江苏省自然科学基金资助(BK2004011)

摘　　要：探讨丙型肝炎病毒(HCV)F基因对PI3K表达的影响。应用PCR技术从含有HCV全长开放阅读框的质粒中获得HCV-F基因片段,利用基因重组技术将其克隆至真核表达载体pcDNA3.0(-)中。通过酶切、PCR及测序鉴定,HCV-F基因已正确插入到pcDNA3.0(-)中,再利用脂质体转染HepG2细胞。经RT-PCR及Westernblot检测,HCV的F基因在HepG2细胞中获得表达,而且在表达重组HCV-F的转染HepG2细胞中,检测到PI3K蛋白的表达。实验表明HCV-F可在体外激活PI3K及其信号通路。To reveal the influence of HCV-F on PI3K, The relationship between F and PI3K in vitro was explored. Full length F gene of HCV was amplified by PCR, using the plasmid containing HCV fulllength open reading frame （ORF） as template, and cloned into the eukaryotic expressing plasmid pcDNA3.0（-） by DNA recombination technique. The recombinant vector was identified by digestion with restriction enzymes and polymerase chain reaction and by direct sequencing. Then both the recombinant vector pcDNA3. 0-F and the control vector pcDNA3. 0 （-） were transfected HepG2 cell using Lipofectamin2000. Expressing of F gene was proved by RT-PCR and Western blot analysis in transfected HepG2 cell. The PI3K protein expressed in the HepG2 cell expressing recombinant HCV-F was further examined. HCV-F can activate PI3K in vitro and its signal cascade pathway.

关 键 词：丙型肝炎病毒F蛋白 HEPG2细胞 磷脂酰肌醇-3激酶 

分 类 号：Q78[生物学—分子生物学]