Expression and purification of human S100A2-GST fusion protein in prokaryotic cells

Objective:To obtain human S100A2-GST fusion protein for further research on human S100A2(hS100A2)protein function and its interactions with other proteins and systems.Methods:hS100A2 gene from pHAHA-hS100A2 was subcloned into an expression vector pGST-moluc to construct pGST-moluc-hS100A2.The new plasmid was identified by digestion with XhoⅠand EcoRⅠ.The recombinant BL21 was induced with IPTG to express recombinant fusion protein hS100A2-GST,which was purified by Glutathion-Sepharose 4B ball beads,identified by SDS-PAGE and Western Blot,and quantified by Bradford method.Results:After the digestion,the recombinant plasmid was cutted into two fragments,300 bp and 5kb.After treated with IPTG,the recombinant BL21 strain expressed a protein,which was about 36 kD and was recognized specifically by an-ti-hS100A2.Its yield was 5 mg/L bacterial culture.After isolated by Glutathion-Sepharose 4B ball beads,its purity was 92%.Conclusion:hS100A2-GST expression plasmid pGST-moluc-hS100A2 was constructed successfully.hS100A2-GST fusion pro-tein could be expressed in Escherichia coli with higher yield and isolated with higher purity,which lays the foundation for the follow-up of the hS100A2 research.