Eukaryotic expression of HBV CTL epitope single chain trimer gene and construction of HBsAg-SCT recombinant adenovirus vector

Objective:To construct an eukaryotic expression vector of the HBV CTL antigen epitope single chain trimer(HBsAg-SCT) gene,and express in 293T cell,at the same time,construct the adenoviral expression vector.Methods:An oligonucleotide of HBV CTL epitope was synthesized and fused with H-2Ld DNA molecule to construct the eukaryotic expression vector carrying the HBsAg-SCT gene,then the recombinant plasmid was transfected into 293T cells,the expression was detected by flow cytometry.The HBsAg-SCT gene was subcloned into an adenoviral expression vector,then transfected into 293A cells for packaging and amplification of recombinant adenovirus.Results:The eukaryotic expression vector carrying HBsAg-SCT gene was constructed successfully,and the expression in 293T cell was proved.The high titer recombinant adenovirus was generated utilizing the recombinant adenovirus vector.Conclusion:The recombinant eukaryotic expression vector expressed HBsAg-SCT effectively,the recombinant adenovirus can be used in further study of the immunity therapy for chronic HBV.