Construction of the eukaryotic expression vector of compound CTL multi-epitope gene of HCV in Chinese people and expression of the gene in COS-7 cells

Objective: To construct and identify the eukaryotic expression vector of compound T cell multi-epitope gene of HCV in Chinese people. Methods: Multi-epitope gene of HCV was screened by bioinformatic method and synthesized. The gene was cloned into eukaryotic expression vector pEGFP-N1. Thus, the plasmid pEGFP-mcf was constructed and then transfected into COS-7 cells with liposome. The mRNA expression of multiple-epitope fragment (mcf) was detected by RT-PCR. The expression and subcellular localization of the fusion protein were detected by Western blot and fluorescent microscope. Results: The eukaryotic expression vector of compound T cell multi-epitope gene of HCV was successfully constructed. Both RT-PCR and Western blot verified that the compound T cell multi-epitope gene was expressed in COS-7 cells. The green fluorescence of mcf-EGFP fusion protein was mainly located in the cytoplasm of COS-7 cells, and that of pEGFP protein was in both cytoplasm and nucleus of COS-7 cells. Conclusion: The eukaryotic expression vector of compound T cell multi-epitope gene of Chinese people was constructed and expressed successfully in COS-7 cells, which has laid a foundation for further study on cytoxicity T lymphocyte multi-epitope vaccine.