[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.