[Researches on cloning and expression of the gene encoding Schistosoma japonicum tetraspanins extracellular loop 2 and its immunogenicity].

Sichuan Da Xue Xue Bao Yi Xue Ban

State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, China.

Published: November 2008

Objective: To clone and express the gene (Sj-tsp-2) encoding extracellular loop 2 (EC-2) of tetraspanins (TSPs) of Schistosoma japonicum (Chinese strain), and then to study the antigenicity and immunogenicity of this protein.

Methods: Synthesized Sj-tsp-2 gene was cloned into prokaryotic expression vector pET32a to generate pET32a-Sj-tsp-2 recombinant plasmid, and transformed into competent E. coli BL21 (DE3). After inducing with IPTG, the expressed fusion protein was purified under nondenaturing conditions.

Results: The recombinant was confirmed by sequence analysis. The size of fusion protein and interest peptide was accords with the theoretical value by SDS-PAGE analysis. Additionally, the results of Western Blotting and ELISA demonstrated that the expressed protein had good immunogenicity. More importantly, we confirmed that TSP-2 is mainly located on the surface of S. japonicum.

Conclusion: The successful expression and purification of recombinant protein Trx-Sj-TSP-2 will be very helpful for the further study of its protection role in animals.

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