嵌合中国分离株基因的丙型肝炎病毒(1b/2a)细胞培养模型的建立  

Development of Novel Chimeric Hepatitis C Virus Cell Culture Systems Derived from Intergenotypic 1b/2a(Chinese Isolates/JFH1) Recombinant Genome

作　　者：卢莎[1,2] 邓瑶[2] 沈晓玲[1] 管洁[2] 周为民[2] 王岳[2,3] 阮力[2] 谭文杰[2,3] 

机构地区：[1]内蒙古医学院微生物教研室,内蒙古呼和浩特010059 [2]中国疾病预防控制中心病毒病预防控制所,北京100052 [3]病毒基因工程国家重点实验室,北京100052

出　　处：《生物技术通讯》2009年第4期502-505,共4页Letters in Biotechnology

基　　金：国家高技术研究发展计划(2007AA02Z455);国家自然科学基金(30571673;30671961)

摘　　要：目的:建立嵌合中国分离株基因的丙型肝炎病毒(HCV)细胞培养模型。方法:利用3片段融合PCR的方法将中国HCV河北分离株(1b)的全长包膜蛋白基因引入JFH1(2a)株基因骨架,构建包膜蛋白基因区相互置换的嵌合HCV(1b/2a)全长基因组,经线性化后体外转录获得全长RNA,转染Huh7.5.1细胞系,用免疫荧光及蛋白印迹实验检测。结果:该RNA可以产生具有体外感染活性的嵌合HCV,且感染性可在共同培养的细胞间传播。结论:首次在国内建立了嵌合中国HCV分离株基因的HCV细胞培养体系。Objective： To development a novel chimeric genotype 1b/2a hepatitis C virus（HCV） cell culture system conraining envelope glycoprotein derived from China patient. Methods： We use overlapping PCR to clone HCV envelope gene （coding aa 170-746） which isolated from Hebei province patient（1b） into JFH1（2a） backbone. A full genome chimeric（1b/ 2a） HCV infectious RNA was synthesized by in vitro transcription. The Huh7.5.1 cells were transfected with this novel chimeric RNA transcripts of an intergenotypic 1b/JFHl（2a） recombinant. The virus cultures were examined by immunostaining and Western blotting. Results： This novel chimeric HCV（1b/2a） virus can be produced in Huh7.5.1 cell by recombinant RNA transfection and the virus was infectious in cell culture and transmissible among the cells. Conclusion： The data suggest that a novel HCV culture system, which based on integrating the envelope glycoprotein gene（1b, Chinese isolate） into JFH1 backbone（2a）, was first reported in China mainland. This chimeric HCV cell culture system provides a new tool for researching HCV and developing antiviral stragegies.

关 键 词：丙型肝炎病毒 嵌合病毒 细胞培养 

分 类 号：Q813.1[生物学—生物工程] R373.21[医药卫生—病原生物学]