擬態(tài)弧菌OmpU蛋白B細(xì)胞表位的篩選鑒定及其免疫活性研究

李槿年

摘 要：擬態(tài)弧菌是一種人和水產(chǎn)動物共患病病原菌，不僅引起水產(chǎn)動物嚴(yán)重的腹水病，而且可經(jīng)水體和水產(chǎn)品感染人類，引起腹瀉和食物中毒。擬態(tài)弧菌OmpU蛋白是一種具有良好抗原性和保守性的首選診斷抗原和疫苗候選分子，但其抗原表位仍不清楚。對此，該項(xiàng)目擬開展“擬態(tài)弧菌OmpU蛋白B細(xì)胞表位及其免疫活性研究”。應(yīng)用生物信息軟件預(yù)測OmpU蛋白B細(xì)胞線性表位，進(jìn)而利用合成肽ELISA鑒定預(yù)測表位，確定優(yōu)勢B細(xì)胞線性表位。再以O(shè)mpU多克隆抗體和單克隆抗體為靶標(biāo)從噬菌體隨機(jī)肽庫中篩選鑒定優(yōu)勢B細(xì)胞模擬表位。最后，對表（擬）位噬菌體克隆進(jìn)行免疫活性檢測，分析不同表位的免疫活性差異。本項(xiàng)目預(yù)期研究成果不僅為建立以表位為基礎(chǔ)的特異快速的診斷方法和研制表位疫苗奠定基礎(chǔ)。

關(guān)鍵詞：擬態(tài)弧菌 OmpU蛋白 B細(xì)胞表位 免疫活性

Abstract： Vibrio mimicus （V. mimicus） is a zoonisis pathogens. It not only cause ascitic fluid disease of aquatic animals， but also cause human gastroenteritis and food poisoning through contacting aquatic environment and aquatic products. The out membrance protein U （OmpU） of V. mimicus with higher antigenicity and conservative property is taken as a primary diagnostic antigen and good candidate protein for developing vaccine， but B-cell epitopes of OmpU protein are not clear now. In order to design new vaccine and diagnosing reagent in the future， the B-cell eptopes of OmpU protein of V. mimicus and their immune characteristics were researched. Firstly， B-cell epitopes of OmpU protein were analyzed with bio-softwares Biosun and Hopp-wood. The putative epitopes peptides were synthesized and were defined by peptide enzyme-linked immunosorbent assay. Secondly， in order to define B-cell mimotopes of OmpU protein， the phage clones binding to purified polyclonal antibody and monoclonal antibody against OmpU protein were selected from the 12-mer random peptide library. Then positive monoclonal phages were sequenced and analyzed. Finally， the immunogenicity and immunoreactivity of differnt phage clones containing B-cell epitopes were determined with immunoassay to compare their immune characteristics.

Key Words： Vibrio mimicus； OmpU protien； B cells epitopes； Immune characteristics

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