Cloning, expression and protective immunity evaluation of the full-length cDNA encoding succinate dehydrogenase iron-sulfur protein of Schistosoma japonicum |
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Authors: | JunLong Yu ShiPing Wang WenKai Li Gan Dai ShaoRui Xu Zhuo He XianChu Peng SongHua Zhou XueQin Liu |
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Institution: | (1) Department of Parasitology, Xiangya School of Medicine, Central South University, Changsha, 410078, China |
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Abstract: | 1071-bp fragment was obtained from the Schistosoma japonicum (Chinese strain) adult cDNA library after the 3′ and 5′ ends of the incomplete expression sequence tag (EST) of succinate
dehydrogenase iron-sulfur protein of Schistosoma japonicum (SjSDISP) were amplified by the anchored PCR with 2 pairs of primers designed according to the EST of SjSDISP and the sequence of multiclone sites of the library vector. Sequence analysis indicated that the fragment was a full-length
cDNA with a complete open reading frame (ORF), encoding 278 amino acid residues. The fragment was cloned into prokaryotic
expression vector pQE30, and subsequently sequenced and expressed in Escherichia coli. SDS-PAGE and Western-blot analyses showed that the recombinant protein was about 32 kD and could be recognized by the polyclonal
antisera from rabbits immunized with Schistosoma japonicum adult worm antigen. Compared with the FCA controls, mice vaccinated with rSjSDISP (test) or rSjGST (positive control) all revealed high levels of specific antibody and significant reduction in worm burden, liver eggs
per gram (LEPG), fecal eggs per gram (FEPG) and intrauterine eggs. These results suggest that SjSDISP may be a novel and partially protective vaccine candidate against schistosomiasis. In contrast to the worm burden reduction
rate, the higher degree of egg reduction rate in the test group also suggested that SjSDISP vaccine may primarily play a role in anti-embryonation or anti-fecundity immunity.
Supported by the China National Tenth “Five-year Plan” Important Special Program (Grant No. 2002AA2Z3343), the China “863”
Important Program (Grant No. 2004AA2Z3530), Hunan Province Eleventh “Five-year Plan” Improtant Special Program (Grant No.
2006SK1001), and Hunan Province Tenth “Five-year Key Xueke Plan” (Grant No. 03-985-3-7) |
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Keywords: | Schistosoma japonicum iron-sulfur protein of succinate dehydrogenase cloning expression protective immunity |
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