PROKARYOTIC EXPRESSION OF Sj16 GENE OF SCHISTOSOMA JAPONICUM CHINESE STRAIN

Objective To further study the immunodulating function of Schistosoma japonicum Sj16. Methods A couple of primers were designed according to the known sequence of Schistosoma mansoni Sm16 and the Sj16 gene was obtained by amplification from cDNA library of Schistosoma japonicum Chinese strain by using PCR technique. By cloning Sj16 gene into a prokaryotic expression vector, pGEX-4T-1-Sj16, a recombinant pGEX-4T-1-Sj16 was constructed and transferred into E.coli BL21/DE3. The positive recombinant pGEX-4T-1- Sj16 was screened and identified by agarose gel electrophoresis, endonuclease digestion and PCR technique. The BL21/DE3 which was transformed with pGEX-4T-1- Sj16 was induced by IPTG, and the expressed protein was analyzed by SDS-PAGE and Western-blot. Results The specific fragment of Sj16 was amplified and the recombinant prokaryotic expression plasmid pGEX-4T-1- Sj16 was successfully constructed. After being induced by IPTG, the transformed BL21/DE3 expressed an about 40 kDa protein which was fused with GST. This protein could be recognized by anti-GST antibody in Western-blot test. Conclusion The recombination prokaryotic expression plasmid pGEX-4T-1- Sj16 was constructed and the Sj16 gene was successfully expressed in BL21/DE3, which paved the way for further study on immunomodulatory function of Schistosoma japonicum Sj16 and obtained the knowledge of the immune evasion of schistosome.