Characterisation of a Babesia orientalis apical membrane antigen, and comparison of its orthologues among selected apicomplexans
Characterisation of a Babesia orientalis apical membrane antigen, and comparison of its orthologues among selected apicomplexans
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东方巴贝虫顶膜抗原的表征及其在选定顶复门中的直系同源物的比较
DOI：
10.1016/j.ttbdis.2015.01.010
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发表时间：
2015-01-01
影响因子：
3.2
通讯作者：
Zhao, Junlong
中科院分区：
文献类型：
--
作者：
He, Lan;Fan, Lizhe;Zhao, Junlong
在本研究中，我们确定和鉴定了东方Babelioralis顶端膜抗原1（命名为Bo-ama 1）的完整编码序列;它的长度为1803 bp，编码601个氨基酸（aa）的多肽。Bo-ama-1基因产物（Bo-AMA 1）的大小预计为67 kDa，并含有信号肽。预测成熟的Bo-AMA 1具有一个跨膜区和一个短的胞质尾区（C-末端结构域）。Bo-AMA 1的胞外部分具有三个功能结构域（DI、DII和DIII），具有14个保守的半胱氨酸残基。将含有所有这三个结构域的Bo-AMA 1片段（命名为Bo-AMA 1-DI/II/III）克隆到质粒载体pET-28 a中，并表达为53 kDa的重组（His-融合）蛋白。血清中的抗体来自一个B。东方蜱感染的布法罗在免疫印迹分析中特异性识别该蛋白。针对重组蛋白产生的兔抗体能够检测来自B的红细胞的天然Bo-AMA 1（67 kDa）。感染东方蜱的布法罗。Bo-AMA 1是AMA 1家族的新成员，可能是检测B抗体的良好抗原。东方感染牛。这种蛋白质可能在宿主细胞粘附和B入侵过程中发挥关键作用。东方疟原虫，如在其他生物体如恶性疟原虫和刚地弓形虫中报道的AMA 1。需要进一步的研究来探索这种蛋白的生物学功能，并确定其免疫是否能诱导布法罗对B的保护作用。东方感染(C)2015 Elsevier GmbH. All rights reserved.
In the present study, we identified and characterised the complete coding sequence of Babesia orientalis apical membrane antigen 1 (designated Bo-ama1); it is 1803 bp in length and encodes a polypeptide of 601 amino acids (aa). The Bo-ama-1 gene product (Bo-AMA1) is predicted to be 67 kDa in size and contains a signal peptide. Mature Bo-AMA1 is predicted to have one transmembrane region and a short cytoplasmic tail (C-terminal domain). The extracellular part of Bo-AMA1 has three functional domains (DI, DII and DIII) with 14 conserved cysteine residues. A Bo-AMA1 fragment containing all three of these domains (designated Bo-AMAl-DI/II/III) was cloned into the plasmid vector pET-28a and expressed as a recombinant (His-fusion) protein of 53 kDa. Antibodies in the serum from a B. orientalis-infected water buffalo specifically recognised this protein in immunoblotting analysis. Rabbit antibodies raised against the recombinant protein were able to detect native Bo-AMA1 (67 kDa) from erythrocytes of B. orientalis-infected water buffalo. Bo-AMA1 is a new member of the AMA1 family and might be a good antigen for the specific detection of antibodies produced in B. orientalis infected cattle. This protein is likely to play critical roles during host cell adherence and invasion by B. orientalis, as the AMA1s reported in other organisms such as Plasmodium falciparum and Toxoplasma gondii. Further research is required to explore the biological functions of this protein and to determine whether its immunisation can induce protective effects in water buffalo against B. orientalis infection. (C) 2015 Elsevier GmbH. All rights reserved.