关键词: 钩端螺旋体；原核表达；融合蛋白；活性鉴定

Abstract:

To construct the prokaryotic expression system of the  recombinant advantage antigen epitopes of Leptospira,and to gain the high purity expression products of the  recombinant advantage epitope fusion protein of Leptospira and to provide the basis for research on  the  rapid  method to detect Leptospira.Methods The plasmid pET-rLP was constructed by gene recombinant method.The fusion protein with His-tag was efficiently expressed in the form of inclusion body after IPTG induction.The inclusion body was washed by urea,dissolved and purified by Ni2+ affinity chromatography,and it was identified by SDS-PAGE,Western blotting and ELISA methods.Results There was a specific protein band,which identified the high expression of the recombinant advantage epitope fusion protein of Leptospira by SDS-PAGE analysis and Western blotting,at the molecular weight of 20 000.The ELISA results showed specific reactions with Leptospirosis positive sera,but no cross-reaction with the other positive sera sample(salmonella) using the expression protein.Conclusion The prokaryotic expression system of the  recombinant advantage antigen epitopes of Leptospira is constructed successfully,and the recombinant advantage epitope of  fusion protein of Leptospira with high purity  is gained successfully.

Key words: Leptospira;prokaryotic expression;fusion protein;activity idenfification