Cloning, expression and protective immunity evaluation of the full-length cDNA encoding succinate dehydrogenase iron-sulfur protein of Schistosoma japonicum.
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.
Yu
JunLong
J
Department of Parasitology, Xiangya School of Medicine, Central South University, Changsha 410078, China.
Wang
ShiPing
S
Li
WenKai
W
Dai
Gan
G
Xu
ShaoRui
S
He
Zhuo
Z
Peng
XianChu
X
Zhou
SongHua
S
Liu
XueQin
X
D000818:Animals; D003001:Cloning, Molecular; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics; D018509:DNA, Helminth / Q000235:genetics / Q000302:isolation & purification; D005260:Female; D015723:Gene Library; D015801:Helminth Proteins / Q000235:genetics; D006801:Humans; D007506:Iron-Sulfur Proteins / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D010957:Plasmids; D012549:Schistosoma japonicum / Q000201:enzymology* / Q000235:genetics*; D013385:Succinate Dehydrogenase / Q000235:genetics*
D017931:DNA Primers; D018076:DNA, Complementary; D018509:DNA, Helminth; D015801:Helminth Proteins; D007506:Iron-Sulfur Proteins; D013385:Succinate Dehydrogenase