[Construction and expression of the efficient recombinant plasmid pEE14.1-IFN-α expressing human IFN-α].

AIM: To construct the eukaryotic expression plasmid pEE14.1-IFN-α expressing human IFN-α gene,and to detect the expression of the plasmid in eukaryotic cells.METHODS: The human IFN-α gene amplified by PCR and was linked into pCI-GPI,then inserted into the eukaryotic expression vector pEE14.1.The recombinant plasmid was transfected into the 293T cells,the IFN-α expression was detected by ELISA and Western blotting.RESULTS: Enzyme digestion and sequence analysis showed that the bicistronic eukaryotic expression vector pEE14.1-IFN-α was constructed successfully.The expression of plasmid was detected by ELISA,and the production of IFN-α in supernatant of transfected cells was about 3.15 ng/mL.Also,Western blotting could reveal the characteristic band of IFN-α gene.CONCLUSION: The vector is constructed successfully,which provide a new selection for HBV immunotherapy.