Identification and characterization of a novel 34 kDa merozoite protein in Babesia orientalis
Identification and characterization of a novel 34 kDa merozoite protein in Babesia orientalis
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东方巴贝虫中新型 34 kDa 裂殖子蛋白的鉴定和表征
DOI：
10.1016/j.vetpar.2015.06.031
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发表时间：
2015-09-15
影响因子：
2.6
通讯作者：
Zhao, Junlong
中科院分区：
文献类型：
--
作者：
He, Lan;Fan, Lizhe;Zhao, Junlong
利用B免疫筛选法从东方BabelialiscDNA表达文库中获得了一个新的34 kDa蛋白（BoP 34）。东方蜱感染布法罗血清。BoP 34基因全长1088 bp，包含一个开放阅读框（ORF）、两个非翻译区（UTR）和一个poly（A）尾。该基因的开放阅读框（ORF）长933 bp，编码310个氨基酸，预测分子量为34 kDa。BLAST分析表明，BoP 34基因的核苷酸序列与牛Babybovis基因XM_001611335的核苷酸序列有71%的相似性，后者编码一个核运动家族蛋白。这表明BoP 34是运动家族蛋白的同源物。BoP 34蛋白的结构分析表明，CS结构域可能与热休克蛋白90的ATP酶结构域相互作用。将截短形式的BoP 34克隆到表达载体pET-32 a中，随后从大肠杆菌Rosetta（TM）（DE 3）pLysS菌株中表达并纯化为Trx融合体（命名为rBoP 34-T）。一个B的血清中的抗体。在免疫印迹分析中，东方尾虫感染的布法罗能够识别该蛋白。针对rBoP 34-T产生的兔抗体可以检测B中的天然BoP 34（34 kDa）。东方蜱感染的布法罗红细胞。提示BoP 34可能是一种较好的特异性检测抗B的诊断抗原。布法罗东方蜱抗体的检测。需要进一步的研究来探索这种分子的生物学功能和诊断潜力。(C)2015 Elsevier B. V.版权所有。
A novel Babesia orientalis 34 kDa protein (designated BoP34) was obtained by immunoscreening of a cDNA expression library using 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 that of the Babesia bovis gene XM_001611335, which encodes a nuclear movement family protein. This suggested that BoP34 is a homologous of the movement family protein. Structural analysis of the BoP34 protein indicated a CS domain which may interact with the ATPase domain of the heat shock protein 90. A truncated version of BoP34 was cloned into the expression vector pET-32a and subsequently expressed and purified from the Escherichia coli Rosetta (TM) (DE3) pLysS stain as a Trx-fusion (designated rBoP34-T). Antibodies in the serum of a B. orientails-infected water buffalo were able to recognize this protein in immune-bloting analysis. Rabbit antibodies raised against rBoP34-T could detecte native BoP34 (34 kDa) in B. orientalis-infected water buffalo erythrocytes. These results suggested that BoP34 might be a good diagnostic antigen for the specific detection of anti-B. orientalis antibody in water buffalo. Further research is required to explore the biological function and diagnostic potential of this molecule. (C) 2015 Elsevier B.V. All rights reserved.