临床肝胆病杂志2011，Vol.27Issue(1)：45-48,4.
丙型肝炎病毒CTL表位基因真核表达载体的构建及稳定转染细胞系的建立
Construction of eukaryotic expression vector of HCV CTL epitopes and establishment of stable transfected CHO cell line
摘要
Abstract
Objective To construct eukaryotic expression vector of hepatitis C virus (HCV) cytotoxic T-lymphocyte (CTL) epitopes, and to establish stable transfected CHO cell-lines.Methods The HCV CTL epitopes of different genotypes and the mouse H2 complex were predicted by bioinformatics, then synthesized and inserted into eukaryotic expression vector pEGFP-N3.The recombinant vector was transfected into CHO cells by lipofectamine 2000.After screening with G418, stably transfected CHO cell line was established.The expression of HCV muiti-epitopes was identified by RT-PCR and western-blot and immunofluorescence.Results The eukaryotic expression vector was constructed successfully.The stable transfected CHO cell line was established.Conclusion The establishment of stable transfected CHO cell line and the expression of the target gene provide solid foundation for further experimental studies.关键词
肝炎病毒属/转染/基因表达/表位,T淋巴细胞/T淋巴细胞,细胞毒性Key words
hepacivirus/ transfection/ gene expression/ epitopes, T-lymphocyte/ T-lymphocytes, cytotoxic分类
医药卫生引用本文复制引用
吕欣,尹文,孙梦宁,黄小军,杨敬,姚敏,薛小平,贾战生,徐志凯..丙型肝炎病毒CTL表位基因真核表达载体的构建及稳定转染细胞系的建立[J].临床肝胆病杂志,2011,27(1):45-48,4.基金项目
国家"863"专项课题(2007AA02Z441) （2007AA02Z441）
国家"十一五"重大科技专项(2000ZX10004-715) （2000ZX10004-715）
