[Cloning and expressing of cyclophilin B gene from Schistosoma japonnicum and the analysis of immunoprotective effect].
The present study was intend to clone and express the cDNA encoding Cyclophilin B (CyPB) of Schistosoma japonicum, its preliminary biological function and further immunoprotective effect against schistosome infection in mice. RT-PCR technique was applied to amplify a full-length cDNA encoding protein Cyclophilin B (Sj CyPB) from schistosomula cDNA. The expression profiles of Sj CyPB were determined by Real-time PCR using the template cDNAs isolated from 7, 13, 18, 23, 32 and 42 days parasites. The cDNA containing the Open Reading Frame of CyPB was then subcloned into a pGEX-6P-1 vector and transformed into competent Escherichia coli BL21 for expressing. The recombinant protein was renaturated, purified and its antigenicity were detected by Western blotting, and the immunoprotective effect induced by recombinant Sj CyPB was evaluated in Balb/C mice. The cDNA containing the ORF of Sj CyPB was cloned with the length of 672 base pairs, encoding 223 amino acids. Real-time PCR analysis revealed that the gene had the highest expression in 18-day schistosomula, suggesting that Sj CyPB was schistosomula differentially expressed gene. The recombinant protein showed a good antigenicity detected by Western blotting. Animal experiment indicated that the vaccination of recombinant CyPB protein in mice led to 31.5% worm and 41.01% liver egg burden reduction, respectively, compared with those of the control. A full-length cDNA differentially expressed in schistosomula was obtained. The recombinant Sj CyPB protein could induce partial protection against schistosome infection.
Key Laboratory of Animal Parasitology, Ministry of Agriculture, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Science, Shanghai 200241, China.
Jinbiao
J
Peng
Hongxiao
H
Han
Yang
Y
Hong
Yan
Y
Wang
Fanji
F
Guo
Yaojun
Y
Shi
Zhiqiang
Z
Fu
Jinming
J
Liu
Guofeng
G
Cheng
Jiaojiao
J
Lin
D000818:Animals; D000947:Antigens, Helminth / Q000276:immunology; D003001:Cloning, Molecular; D021983:Cyclophilins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology*; D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics; D007114:Immunization; D051379:Mice; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D012549:Schistosoma japonicum / Q000235:genetics* / Q000276:immunology; D012554:Schistosomiasis japonica / Q000517:prevention & control*; D014614:Vaccines, Synthetic / Q000096:biosynthesis / Q000276:immunology
D000947:Antigens, Helminth; D018076:DNA, Complementary; D011994:Recombinant Proteins; D014614:Vaccines, Synthetic; C081786:cyclophilin B; D021983:Cyclophilins