【摘 要】
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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 n
【机 构】
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State Key Laboratory of Agricultural Microbiology,College of Veterinary Medicine,Huazhong Agricultur
【出 处】
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中国畜牧兽医学会兽医寄生虫学分会第十三次学术研讨会
论文部分内容阅读
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-32a,and subsequently expressed in E.coli RosettaTM(DE3)pLysS stain as a Trx-fusion protein(designated rBoP34-T).Antibodies in the serum of a B.orientalis-infected water buffalo specifically recognized this protein in immunoblot analysis.The rabbit antibodies raised against the rBoP34-T detected native BoP34(34 kDa)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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