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棒状体颈部2（RON2）蛋白的分子克隆与特性分析

Molecular Cloning and Characterization of Rhoptry Neck 2 RON2 Protein.

作者信息

Malobi Ngabu, He Lan, Yu Long, He Pei, He Junwei, Sun Yali, Huang Yuan, Zhao Junlong

机构信息

State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, Hubei 430070, China.

Key Laboratory of Animal Epidemical Disease and Infectious Zoonoses, Ministry of Agriculture, Huazhong Agricultural University, Wuhan, China.

出版信息

J Parasitol Res. 2017;2017:7259630. doi: 10.1155/2017/7259630. Epub 2017 Jul 9.

DOI:10.1155/2017/7259630
PMID:28775897
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5523350/
Abstract

Babesiosis caused by is one of the most prevalent infections of water buffalo transmitted by causing a parasitic and hemolytic disease. The organelles proteins localized in apical membrane especially rhoptries neck and microneme protein form a complex called moving junction important during invasion process of parasites belonging to apicomplexan group, including species. A truncated fragment coding a 936 bps fragment was cloned in pMD-19T and subcloned into pET32 (a) expression vector, expressed in BL21. Purified recombinant RON2 was used to produce polyclonal antibody against RON2. Here, we identified the full sequence of gene encoding the rhoptry neck 2 protein that we named RON2 which is 4035 bp in full-length open reading frame without introns, encoding a polypeptide of 1345 amino acids. Western blot of rRON2 probed with buffalo positive serum analysis revealed a band of around 150 kDa in parasite lysates, suggesting an active involvement during invasion process. These findings most likely are constructive in perspective of ongoing research focused particularly on water buffalo babesiosis prevention and therapeutics and globally provide new information for genes comparative analysis.

摘要

由[病原体名称未给出]引起的巴贝斯虫病是水牛最常见的感染之一，通过[传播途径未给出]传播，导致一种寄生性和溶血性疾病。定位于顶膜的细胞器蛋白，特别是棒状体颈部和微线体蛋白形成一种称为移动连接的复合物，在属于顶复门的寄生虫（包括[相关物种未给出]物种）的入侵过程中起重要作用。编码936 bps片段的截短片段被克隆到pMD-19T中，并亚克隆到pET32(a)表达载体中，在BL21中表达。纯化的重组RON2用于制备抗RON2的多克隆抗体。在这里，我们鉴定了编码棒状体颈部2蛋白的基因的完整序列，我们将其命名为RON2，其全长开放阅读框为4035 bp，无内含子，编码一个1345个氨基酸的多肽。用布氏阳性血清分析对rRON2进行蛋白质免疫印迹，在寄生虫裂解物中显示出一条约150 kDa的条带，表明在入侵过程中发挥了积极作用。这些发现从正在进行的特别是针对水牛巴贝斯虫病预防和治疗的研究角度来看很可能具有建设性，并为全球基因比较分析提供了新信息。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6226/5523350/8dddb020759a/JPR2017-7259630.006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6226/5523350/12b73de6141b/JPR2017-7259630.001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6226/5523350/7f07a0469682/JPR2017-7259630.002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6226/5523350/9e1f9d7e4ba6/JPR2017-7259630.003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6226/5523350/d3d75c6d585a/JPR2017-7259630.004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6226/5523350/0dbdaac8c430/JPR2017-7259630.005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6226/5523350/8dddb020759a/JPR2017-7259630.006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6226/5523350/12b73de6141b/JPR2017-7259630.001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6226/5523350/7f07a0469682/JPR2017-7259630.002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6226/5523350/9e1f9d7e4ba6/JPR2017-7259630.003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6226/5523350/d3d75c6d585a/JPR2017-7259630.004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6226/5523350/0dbdaac8c430/JPR2017-7259630.005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6226/5523350/8dddb020759a/JPR2017-7259630.006.jpg

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Infect Immun. 2016 Apr 22;84(5):1574-1584. doi: 10.1128/IAI.00107-16. Print 2016 May.
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Parasit Vectors. 2015 Oct 16;8:543. doi: 10.1186/s13071-015-1158-x.
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Characterisation of a Babesia orientalis apical membrane antigen, and comparison of its orthologues among selected apicomplexans.
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