Molecular cloning and functional characterization of Schistosoma japonicum enolase which is highly expressed at the schistosomulum stage.
Enolase is a key enzyme in the glycolytic pathway; recent studies have also shown that enolase is found on the surface of several parasites, where it acts as a plasminogen-binding protein. In the present study, the enolase of Schistosoma japonicum has been cloned and expressed. In western blot analysis, the recombinant enolase from S. japonicum ( rSjENO) was recognized by rabbit sera directed against an antigen preparation from adult worms. Kinetic measurement revealed that rSjENO possesses good enzymatic activity. The real-time PCR showed that the enolase gene was highly expressed at 18-28 days of the life cycle. Immunofluorescence testing showed that SjENO was located mainly on the surface as well as in the inner tissues of the worms. Ligand-blotting analysis indicated that rSjENO could bind to human plasminogen as its receptor. In addition, a 24.28% reduction in the liver egg count and a reduction of 21.45% in the fecal egg count were observed in BALB/c mice vaccinated with rSjENO when compared with blank control mice. An ELISA assay suggested that high levels of specific IgG antibody could be induced by rSjENO in vaccinated mice.
Yang
Jianmei
J
Shanghai Institute of Veterinary Research, Chinese Academy of Agricultural Sciences, Key Laboratory of Animal Parasitology, Ministry of Agriculture of China, Shanghai, People's Republic of China.
Qiu
Chunhui
C
Xia
Yanxun
Y
Yao
Lixiao
L
Fu
Zhiqiang
Z
Yuan
Chunxiu
C
Feng
Xingang
X
Lin
Jiaojiao
J
D000595:Amino Acid Sequence; D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D018507:Gene Expression Regulation, Developmental; D015801:Helminth Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D051379:Mice; D008807:Mice, Inbred BALB C; D010270:Parasite Egg Count; D010751:Phosphopyruvate Hydratase / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D011817:Rabbits; D012549:Schistosoma japonicum / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism*; D012554:Schistosomiasis japonica / Q000469:parasitology; D017422:Sequence Analysis, DNA; D014611:Vaccination
D000909:Antibodies, Helminth; D015801:Helminth Proteins; D010751:Phosphopyruvate Hydratase