Construction, expression and Purification of a Novel Human Hepatic Growth Factor: Hepatopoietin Cn and Research on its Biological Activity in vitro
新的肝细胞生长因子HPPCn的表达、纯化及体外活性检测

Objective:To construct,express and purify recombinant human HPPCn protein and detect its biological acitivity in vitro.Methods:The prokaryotic expression vector pET-24a( )-HPPCn was constucted by template pUC18-HPPCn,and then was transformed into E.coli BL21,inducing it with 0.4mmol/L IPTG for 4 hours.The expression and solubility of recombinant protein was detected by SDS-PAGE.The interest protein was purified by anion exchange chromatography and its immunogenicity was identitied by Western blot analysis.Its biological activity in vitro was detected by MTT and 3H-TdR assays.Results:The rhHPPCn was highly expressed after IPTG inducing,and 23% of HPPCn was resoluble in thalline.The purity of HPPCn protein reached over 94% after twice anion exchange chromatography,and the Western blot analysis showed it could bind specifically to rabbit-HPPCn-polyclonal antibody.The MTT and 3H-TdR assays showed that the recombinant protein could remarkably promote proliferation of SMMC7721.Conclusion:rhHPPCn is successfully constructed and expressed in E.coli.The interest protein is successfully purified by twice anion exchange chromatography and is detected to promote the proliferation of SMMC7721.