Study on prokaryotic expression of hantaan virus S and M chimeric gene segment

Aim To obtain fused expression of hantaan virus glycoprotein G1 and nucleoprotein fragment including major antigen sites in E. coli .Methods The prokaryotic fusion expression vector pGEX 4T2 S 0.7G1 was constructed through connection of 0.7 kb segment of 5′terminal in S gene region with G1 gene segment encoded by M gene from hantaan virus 76 118 strain and cloned into pGEX 4T2. The expression of the fusion protein was induced in E.coli XL1 Blue, and then the expression product was analyzed by ELISA and Western blot. Results It was proved that the prokaryotic fusion expression vector pGEX 4T2 S 0.7G1 was constructed successfully by restriction enzyme analysis. After IPTG induction, ELISA results showed that the fusion protein could bind specifically to the hantavirus nucleoprotein specific mAb, and Western blot result exhibited a novel protein band with M r more than 1×10 5 which was encoded by S 0.7G1 and GST gene. Besides, a series of protein bands with different molecular weights which could bind to the hantavirus nucleoprotein specific mAb was detected. Conclusion The fusion protein GST S 0.7G1 with specific binding activity to nucleoprotein is obtained. It lays the foundation for the further research on genetic engeneering vaccine for hantaan virus.