Construction of eukaryotic expression vector of HCV CTL epitopes and establishment of stable transfected CHO cell line / 临床肝胆病杂志

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 multi-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.