Construction of Prokaryotic Expression Vector Carrying HBx-d382 and HBx-d431 Genes from Hepatocellular Carcinoma Tissue

Objective To construct the prokaryotic expression vector carrying HBx-d382 and HBx-d431 genes for exploring the relationship between HBx deletion mutant and hepatocellular carcinoma.Methods HBx-d382 and HBx-d431 genes with EcoRⅠ and XhoⅠ endoenzyme sites were obtained by using PCR.The genes were cloned into pGM-T and produced pGM-T / HBx-d382 and pGM-T / HBx-d431 plasmids.The plasmids were transferred to E.coli BL21(DE3).Recombinant plasmid sequence were subsequently verified by EcoRⅠ and XhoⅠ endoenzyme digestion and auto-sequencing assay.Results We have constructed the prokaryotic expression vector of HBx-d382 and HBx-d431 carrying the integrated HBx-d382 or HBx-d431 gene fragment.Conclusion pET-28a(+) / HBx-d382 and pET-28a(+) / HBx-d431 prokaryotic expression vectors can be used for the study of HBx-d382 and HBx-d431 proteins.