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过表达、纯化及对嗜水气单胞菌 AL09-73 鞭毛蛋白的免疫反应。

Over-expression, purification and immune responses to Aeromonas hydrophila AL09-73 flagellar proteins.

机构信息

United States Department of Agriculture, Agricultural Research Service, Aquatic Animal Health Research Unit, 990 Wire Road, Auburn, AL 36832-4352, USA.

出版信息

Fish Shellfish Immunol. 2011 Dec;31(6):1278-83. doi: 10.1016/j.fsi.2011.09.016. Epub 2011 Sep 25.

Abstract

Aeromonas hydrophila is ubiquitous in aquatic environments worldwide and causes many diseases in fish as well as human. Recent outbreaks of aeromonad diseases in channel catfish prompted us to investigate catfish immune responses during infection of A. hydrophila. In this communication, we report to amplify, over-express, purify and characterize 19 A. hydrophila flagellar proteins. All recombinant proteins were confirmed by nucleotide sequencing of expression plasmids, SDS-PAGE analysis and His tag Western blot of induced proteins. Our preliminary result also showed that the purified recombinant FlgK protein reacted strongly to sera from experimentally infected catfish, suggesting that this protein has potential for a novel target for vaccine development. It is also anticipated that these recombinant proteins will provide us with very useful tools to investigate host immune response to this microorganism.

摘要

嗜水气单胞菌在全球水生环境中无处不在,可引起鱼类和人类的多种疾病。最近,斑点叉尾鮰暴发了气单胞菌病,促使我们研究感染嗜水气单胞菌时斑点叉尾鮰的免疫反应。在本通讯中,我们报告了对 19 种嗜水气单胞菌鞭毛蛋白进行扩增、过表达、纯化和特性鉴定。所有重组蛋白均通过表达质粒的核苷酸测序、SDS-PAGE 分析和诱导蛋白的 His 标签 Western blot 进行确认。我们的初步结果还表明,纯化的重组 FlgK 蛋白与经实验感染的斑点叉尾鮰血清强烈反应,这表明该蛋白可能成为疫苗开发的新靶标。预计这些重组蛋白还将为我们研究宿主对这种微生物的免疫反应提供非常有用的工具。

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