[Fusion expression and antigenicity analysis of miracidial antigen from eggs of Schistosoma japonicum].
OBJECTIVE
To express the miracidial antigen from eggs of Schistosoma japonicum (Chinese mainland strain) (SjMP10), and investigate the role of the miracidial antigen during the hepatic granuloma formation of schistosomiasis.


METHODS
A pair of specific primers was designed and synthesized according to the nucleotide sequence of the open reading frame of the miracidial antigen gene. The open reading frame of the miracidial antigen gene was amplified, digested by restrictive enzyme (BamHI, SolI), and cloned directly into the expression plasmid pGEX-4T-3 to construct the recombinant plasmid. The recombinant plasmids were transformed into E. coli BL21(DE3), and induced by IPTG to express the fusion protein of GST-SjMP10. The expressed fusion protein was purified by electric elution method, and its antigenicity was examined by Western blotting and lymphocyte proliferation test.


RESULTS
The gene of miracidial antigen was cloned into the expression plasmid pGEX-4T-3. After induced by IPTG, the recombinant expressed a fusion protein of GST-SjMP10, with a molecular weight of 39 000 approximately. The purified fusion protein showed proper antigenicity that could be recognized by the sera of rabbits heavily infected by Schistosoma japonicum and could stimulate the proliferation of splenic lymphocytes of infected BALB/c mice.


CONCLUSIONS
The miracidial antigen from eggs of Schistosoma japonicum was expressed successfully.
Liu
Jian-Fa
JF
School of Medicine, Ningbo University, Ningbo 315211, China.
Yu
Chuan-Xin
CX
Zhu
Yin-Chang
YC
Yin
Xu-Ren
XR
Xu
Yong-Liang
YL
D000818:Animals; D000947:Antigens, Helminth / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D015153:Blotting, Western; D005982:Glutathione Transferase / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D010063:Ovum; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000276:immunology; D012549:Schistosoma japonicum / Q000276:immunology*; D013154:Spleen / Q000276:immunology
D000947:Antigens, Helminth; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase