Adr亚型乙型肝炎病毒转染细胞模型的构建  

Construction of cell models transfected with HBV adr subtype

作　　者：赵艳芳[1] 闫永平[1] 张磊[1] 王安辉[1] 苏海霞[1] 门可[1] 张景霞[1] 徐德忠[1] 

机构地区：[1]第四军医大学军事预防医学系流行病学教研室,西安710032

出　　处：《中国公共卫生》2006年第9期1066-1068,共3页Chinese Journal of Public Health

基　　金：国家自然科学基金重点课题(30230320)

摘　　要：目的建立乙型肝炎病毒(HBV)复制状态的细胞模型。方法利用分子亚克隆技术,将9600bp的adr亚型HBV全基因克隆至pcDNA3的EcoRI和HindIII位点构建pcDNA3-3HBV,通过脂质体介导的方法转染HepG2细胞,G418筛选。结果成功构建了质粒pcDNA3-3HBV,稳定转染后培养上清,ELISA检测结果显示,HBsAg、HBeAg阳性,且PCR检测前S/S基因阳性。PCR证实转染细胞中有HBV cccDNA存在,RT-PCR证实HBV S基因mRNA的表达。结论重组质粒pcDNA3-3HBV能在HepG2细胞中表达、转录、复制。Objective To establish HBV replication cell models. Methods Using molecular subclone technique, pcDNA3-3HBV was constructed by insertion of 9 600 bp fragment of HBV adr subtype genome into the EcoRI and HindⅢ sites of pcDNA3. HepG2 was transfected with pcDNA3-3HBV by using lipofectamine transfection reagent and screened with antibiotic G418. Results The plasmid pcDNA3-3HBV was constructed successfully. After stable transfection, HBsAg and HBeAg were positive in the supernatant which tested by ELISA. S gene and pre-S gene were also positive which tested by PCR. cccDNA was found in the cells by PCR, and HBV pre-S1 mRNA was measured by RT-PCR. Conclusion pcDNA3- 3HBV can be expressed, transcribed and replicated in HepG2 cells.

关 键 词：乙型肝炎病毒 ADR亚型 真核表达质粒 转染 基因表达 

分 类 号：R373.2[医药卫生—病原生物学]