Abstract: Constructing prokaryotic expression vector of Eg95 gene from Echinococcus granulosus and expressing fusion protein in E. coli BL21 (DE3) followed by gained purified protein.Eg95 gene was subcloned into pET-44a (+) from clone vector pMD19-T-Eg95 to construct the prokaryotic expression vector pET-Eg95. Nus-Eg95 fusion protein was expressed successfully in E. coli BL21(DE3) at an optimized expression condition and then was purified after identification by SDS-PAGE and western blot. Prokaryotic expression vector pET-Eg95 expressed efficiently in E. coli BL21(DE3) . A high expression level was achieved by inducing the bacteria with 0.1 mmol /L IPTG，at 0.6 of OD600 value for 5 h at 37℃. Purified soluble fusion protein was identified to be the correct target protein by SDS-PAGE and western blot with favorable biological activity. The prokaryotic expression vector pET-Eg95 was constructed successfully and expressed at a high level under the optimized expression condition. The fusion protein can be used to immunoblot assay as a specific antigen react with the antibodies in serum.

Key words: Echinococcus granulosus, Eg95, Prokaryotic expression, Protein purification, Biological activity

CLC Number: 

• S826

• S852.7