Construction of GST/HIF-1α fusion protein expression vectors and the expressions in Escherichia coli.

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-1α(HIF-1α)and its deletion fragments were amplified from the plasmid pcD-NA3.1-HIF-1α by PCR and inserted into pGEX-4T-2 by BamHI and Not I. The positive recombinants 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-1α and its deletion mutants were successfully constructed and confirmed by enzyme digestion and sequencing. The desired GST/HIF-1α fusion proteins were expressed and confirmed by Western blot. Conclusion The prokaryotic expression plasmid of HIF-1α and its deletion mutants were successfully constructed and the expression of fusion proteins was confirmed. This study provides the basis for the further research on purifying HIF-1α protein and the biological function of HIF-1α.