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


CONCLUSIONS
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.
Fu
Yi-jie
YJ
State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, China.
Yang
Li
L
Liu
Kang
K
Yuan
Chuang
C
Chen
Xian-cheng
XC
Wei
Yu-quan
YQ
D000818:Animals; D000947:Antigens, Helminth / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors; D015801:Helminth Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D007114:Immunization; D008565:Membrane Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D051379:Mice; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D012549:Schistosoma japonicum / Q000235:genetics* / Q000276:immunology / Q000378:metabolism
D000947:Antigens, Helminth; D015801:Helminth Proteins; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins