Construction of GST/HIF-1α Fusion Protein Expression Vectors and the Expressions in Escherichia coil / 中国医科大学学报

ABSTRACT

Objective To construct GST/HIF1α fusion protein expression vector and induce its expression in Escherichia coli (E.coli). Methods The coding sequence of hypoxia inducible factor-la (HIF-la) and its deletion fragments were amplified from the plasmid pcD-NA3.1-HIF-la by PCR and inserted into pGEX-4T-2 by BamHI and Not I. The positive recombinanls were identified by restriction endonu-clease digestion and DNA sequencing. Then they were transformed into E.coli BL21, induced by IPTG and identified by SDS-PAGE and Western blot. Results The prokaryotic expression plasmid pGEX-4T-2-HIF-lα and its deletion mutants were successfully constructed and confirmed by enzyme digestion and sequencing. The desired CST/HIF-1α fusion proteins were expressed and confirmed by Western blot. Conclusion The prokaryotic expression plasmid of HIF-la and its deletion mutants were successfully constructed and the expression of fu-sion proteins was confirmed. This study provides the basis for the further research on purifying HIF-la protein and the biological function of HIF-lα.