猪圆环病毒2型Cap蛋白B细胞表位的预测及鉴定

发布时间：2017-12-26 21:23

本文关键词：猪圆环病毒2型Cap蛋白B细胞表位的预测及鉴定　出处：《郑州大学学报(理学版)》2017年03期 　论文类型：期刊论文

更多相关文章： 猪圆环病毒型 Cap蛋白 原核表达 表位

【摘要】：利用生物信息学技术和原核表达系统,筛选猪圆环病毒2型(PCV2)核衣壳蛋白(Cap)的B细胞表位,为进一步揭示PCV2的免疫调控机制提供研究依据.运用生物信息学软件预测PCV2 Cap蛋白的亲水性、表面可及性、蛋白柔韧性、抗原性和B细胞线性表位;根据预测结果,初步确定Cap蛋白潜在的B细胞线性表位主要位于58~66、79~91、151~162、174~189、204~213和221~233位氨基酸.设计特异性引物,扩增Cap蛋白潜在表位区域基因,并克隆到p ET32a(+)原核表达载体上,转化至表达菌BL21(DE3)plys S中,在IPTG诱导下表达目的蛋白;利用PCV2阳性血清鉴定Cap蛋白的B细胞线性表位,筛选出3个能与血清结合的肽段,即Cap77-95、Cap174-189和Cap221-233;对获得的肽段进行在线同源比对,发现3个片段均为PCV2 Cap蛋白的特有序列,有可能成为PCV2的特异性表位.
[Abstract]:Using bioinformatics technique and the prokaryotic expression system, screening of porcine circovirus type 2 (PCV2) nucleocapsid protein (Cap) of the B cell epitope, provide a theoretical basis for the further exploration of the mechanism of immune regulation of PCV2. By using bioinformatics software PCV2 Cap protein predicted hydrophilicity, surface accessibility, flexibility, protein the antigenicity and the linear B cell epitope; according to the forecast results, preliminary determination of Cap protein B cell linear potential epitopes are mainly located in 58~66, 79~91, 151~162, 174~189, 204~213 and 221~233 amino acids. The specific primers were designed, Cap protein gene amplification in latent epitope regions, and cloned into P (+ ET32a) original the eukaryotic expression vector and transformed into E.coli BL21 (DE3) plys S in protein expression induced by IPTG; using the linear B cell epitopes of PCV2 Cap protein positive sera were screened, 3 with serum binding peptides, Cap77-95, C Ap174-189 and Cap221-233; online homology comparison of the obtained peptides revealed that all 3 fragments were specific sequences of PCV2 Cap protein, and might become specific epitope of PCV2.
【作者单位】：郑州大学生命科学学院;
【基金】：国家重点研发计划项目(2016YFD0500704) 郑州市科技创新团队项目(131PCXTD622)
【分类号】：S852.651
【正文快照】： 0引言猪圆环病毒(porcine circovirus,PCV)是单链环状无囊膜DNA病毒,该病毒粒子呈二十面体对称结构[1],是已知的最小动物病毒之一[2].根据其基因组成、抗原性和致病性的不同,将其划分成PCV1和PCV2两个类型,其中PCV2是断奶仔猪多系统衰竭综合征(post-weaning multisystemic was

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