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Identification and characterization of a novel 34 kDa merozoite protein in Babesia orientalis

Lan He  Lizhe Fan  Qin Liu  Jinfang Hu  Xiaoyan Miao  Yuan Huang  Pei He  Junwei He  Long Yu  Muhammad Kasib Khan  Yanqin Zhou  Junlong Zhao  
【摘要】:A novel gene encoding a 34 kDa protein of Babesia orientalis(designated BoP34) was obtained by immunoscreening of a cDNA expression library with B.orientalis infected water buffalo serum.The complete nucleotide sequence of the BoP34 was 1088 bp,which contained one open reading frame(ORF),two untranslated regions(UTRs) and a poly(A) tail.The length of ORF was 933 bp,encoding a polypeptide of 310 aa with a predicted size of 34 kDa.BLAST analysis showed that the nucleotide sequence of BoP34 had 71%similarity with the sequence of Babesia bovis nuclear movement family protein(XM_001611335).This suggested that BoP34 is a homologous of the movement family protein.The structure analysis of BoP34 amino acid sequence showed a CS domain which may interact with the ATPase domain of the heat shock protein 90.Partial sequence of BoP34 was truncated and cloned into the expression vector pET-32 a,and subsequently expressed in E.coli Rosetta~(?)(DE3) pLysS stain as a Trx-fusion protein(designated rBoP34-T).Antibodies in the serum of a B.orientalis-intected water buffalo specifically recognized this protein in immunoblot analysis.The rabbit antibodies raised against the rBoP34-T detected native BoP34(34kDa) in B.orientalis-infected water buffalo erythrocytes.The results suggested that BoP34 might be a potential antigen for the specific detection of anti-B.orientalis antibody in cattle.Further research is required to explore the biological function of this molecule and to establish ELISA for the diagnosis of B.orientalis in cattle.

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