HCV NS5A基因在hepG2细胞中的表达以及对PI3K的影响  

Expression of HCV NS5A gene in HepG2 cell and the effect of the NS5A expression on PI3K in vitro

作　　者：胡学芳[1] 邓小昭[1] 刁振宇[1] 张云[1] 周宗安[1] 刘玉[1] 

机构地区：[1]南京军区军事医学研究所分子生物学研究室,江苏南京210002

出　　处：《细胞与分子免疫学杂志》2007年第12期1133-1135,共3页Chinese Journal of Cellular and Molecular Immunology

基　　金：江苏省自然科学基金;江苏省博士后基金资助项目(BK2005004;BK2004011)

摘　　要：目的:探讨HCV-NS5A对PI3K表达的影响。方法:应用PCR技术从含有HCV全长开放阅读框的质粒中获得NS5A全长基因片段,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,NS5A基因已正确插入到pcDNA3.0(-)中,再利用脂质体转染HepG2细胞。结果:经RT-PCR及Western blot检测,HCV的NS5A基因在HepG2细胞中获得表达,而且在表达重组NS5A的转染HepG2细胞中,检测到PI3K蛋白的表达。结论:NS5A可在体外激活PI3K及其信号通路。AIM： To reveal the influence of HCV-NSSA on PI3K, we probe into the relationship between NSSA and PI3K in vitro. METHODS. Full length NSSA gene of HCV was amplified by PCR, using the plasmid containing HCV full-length 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 directly sequencing. Then both the recombinant vector pcDNA3.0 （-）-NSSA and the control vector pcDNA3.0 （-） were transfected HepG2 cell using Lipofectamin2000. RESULTS. Expressing NSSA was proven by RT-PCR and Western blot analysis in transfected HepG2 cell. Further more we examined the PI3K protein expressed in the HepG2 cell expressing recombinant NSSA. CONCLUSION： NSSA can activate PI3K in vitro and its signal cascade pathway.

关 键 词：HCV NS5A HEPG2细胞 基因表达 磷脂酰肌醇-3-激酶(PI3K) 

分 类 号：R392.11[医药卫生—免疫学]