Molecular characterization of a gene encoding a 29-kDa cytoplasmic protein of Babesia gibsoni and evaluation of its diagnostic potentiality
Molecular characterization of a gene encoding a 29-kDa cytoplasmic protein of Babesia gibsoni and evaluation of its diagnostic potentiality
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DOI：
10.1016/s0166-6851(03)00199-3
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发表时间：
2003-10-01
影响因子：
1.5
通讯作者：
Suzuki, H
中科院分区：
文献类型：
--
作者：
Fukumoto, S;Xuan, X;Suzuki, H
用B法筛选吉氏巴氏巴氏杆菌裂殖子cDNA表达文库。吉布森感染的狗血清克隆了编码29-kDa蛋白的cDNA，命名为P29基因。P29基因全长792 bp。计算机分析表明，P29基因含有一个597 bp的开放阅读框和一个250 bp的内含子，编码蛋白质约23.4 kDa。P29蛋白与弓形虫细胞骨架蛋白IMC 1具有同源性。Southern杂交分析表明，P29基因在B中以单拷贝存在。Gibsoni基因组B的天然P29蛋白。通过用抗重组P29小鼠血清进行蛋白质印迹，鉴定了分子量为29 kDa的gibsoni。激光共聚焦显微镜分析显示P29蛋白定位于B细胞的胞浆中。吉布森裂殖子重组P29蛋白在E. coli作为抗原进行酶联免疫吸附试验（ELISA）。ELISA能区分B型。gibsoni感染的狗血清和B.犬亚种感染的犬血清或正常犬血清。此外，对P29蛋白的抗体反应在实验感染的狗在慢性感染阶段保持，表明重组P29蛋白可能是一个有用的诊断试剂用于检测抗体的B。狗的吉布森(C)2003年由Elsevier B. V.出版
A cDNA expression library prepared from Babesia gibsoni merozoite mRNA was screened with B. gibsoni-infected dog serum. cDNA encoding 29-kDa protein was cloned and designated as the P29 gene. The complete nucleotide sequence of the P29 gene was 792 bp. Computer analysis suggested that the sequence of the P29 gene contained an open reading frame of 597 bp with a coding capacity of approximately 23.4 kDa and a single intron of 250 bp. The P29 protein had homology to Toxoplasma gondii cytoskeletal protein IMC1. Southern blot analysis indicated that the P29 gene was present as a single copy in the B. gibsoni genome. The native P29 protein of B. gibsoni with a molecular mass of 29 kDa was identified by Western blotting with anti-recombinant P29 mouse serum. Confocal laser microscopic analysis showed that the P29 protein was located on the cytoplasma of B. gibsoni merozoites. The recombinant P29 protein expressed in E. coli was used as an antigen in an enzyme-linked immunosorbent assay (ELISA). The ELISA was able to differentiate between B. gibsoni-infected dog serum and B. canis subspecies-infected dog serum or normal dog serum. Furthermore, the antibody response against the P29 protein was maintained during the chronic stage of infection in an experimentally infected dog, indicating that the recombinant P29 protein might be a useful diagnostic reagent for the detection of antibodies to B. gibsoni in dogs. (C) 2003 Published by Elsevier B.V.