Construction Of Recombinant S100B Expressing Fusion Protein And Its Immunocompetence Study

Objective: To construct candidate expressing S100B recombinant fusion protein and analyze the immunogenicity and immunoreactivity.To supply a basis for the study of diagnosis for brain injury.Methods and Results:1 .The S100B gene was amplified by RT-PCR. The genes were inserted into the clone vector. The sequence of S100B gene was determined and analyzed .It was showed that recombinant plasmid of S100B gene was constructed successfully.2. Prokarytic system expressing recombinant MBS fusion proteinThe S100B gene was purified and inserted into expression vector pMAL-c2x, It was showed that the recombinant expression system were constructed successfully, 75 kDa recombinant MBS fusion protein was induced by ITPG. MBS fusion protein could be found by SDS-PAGE, which amounted to 31% of the total bacterial protein.3.We have purified the fusion protein by use of amylose chromatography and the purity of fusion protein was 91%. The result suggested that purified fusion protein have strong immunoreactivity.Conclusions: 1. The S100B are highly conserved genes. 2. The prokaryotic expression system TBl(pMS) is most suitable for S100B protein. 3. Amylose resin affinity chromatography is a kind of highefficiency and quick method for purification of MBS fusion protein.This establishes a basis for developing on the protein.