中国生物工程杂志

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.