利用噬菌体展示肽库筛选鸡传染性支气管炎病毒模拟抗原表位  

Screening of Mimotopes of Avian Infectious Bronchitis Virus from Phage Display Peptide Library

作　　者：安烨[1] 王宏俊[1] 张培君[1] 

机构地区：[1]北京市农林科学院畜牧兽医研究所

出　　处：《生物技术通讯》2007年第6期931-934,共4页Letters in Biotechnology

基　　金：北京市科技新星项目(2005B35)

摘　　要：目的:利用噬菌体展示肽库技术筛选鸡传染性支气管炎病毒(IBV)的模拟抗原表位。方法:用IBV阳性血清纯化IgG作为靶标,对噬菌体展示随机12肽库进行筛选,通过ELISA和竞争抑制ELISA鉴定筛选克隆的结合特性,并对阳性克隆提取ssDNA进行测序分析。结果:3轮生物淘洗后,目标噬菌体得到125倍富集。随机挑选50个克隆进行ELISA和竞争抑制ELISA,其中有12个噬菌体克隆可以与IBV阳性血清高特异性结合。测序分析发现,这12个克隆带有2种氨基酸序列,即KSPKHSSSALHF和SFFQLNLHRPTS,且未发现这2种序列与GenBank中已发表的IBV氨基酸序列有同源性。结论:结果提示,这2个肽可能是IBV抗原的模拟表位。Objective： To obtain the mimotope of avian infectious bronchitis virus（IBV） by phage display peptide library. Methods： IgG of IBV was purified from chicken sera and used as a target to screen the phage display random 12- peptide library. The specific phage clones were analyzed for the binding ability with ELISA and competitive inhibition ELISA. And ssDNA of the positive clones were purified for DNA sequencing. Results： After 3 rounds of biopanning, the enriched ratio of phage was increased to 125 times. 50 phage clones were randomly picked and identified by ELISA and competitive inhibition ELISA. 12 positive clones were obtained. Two kinds of amino acid sequences displayed on 12 positive clones, KSPKHSSSALHF and SFFQLNLHRPTS. Compared with sequences of IBV in GenBank, there were no homologous with two peptides amino acid sequences. Conclusion： The results indicated that the peptides might be mimotopes of IBV.

关 键 词：鸡传染性支气管炎 噬菌体展示肽库 抗原表位 

分 类 号：Q789[生物学—分子生物学] R392[医药卫生—免疫学]