用噬菌体随机肽库分析猪繁殖与呼吸综合征病毒(PRRSV)N蛋白单克隆抗体识别的表位  

Identification of the Epitope of PRRSV N Protein Monoclonal Antibody with Phage Displayed Random Peptide Library

作　　者：王玉娥[1] 杨汉春[1] 郭鑫[1] 陈艳红[1] 查振林[1] 

机构地区：[1]中国农业大学农业部预防兽医学重点开放实验室,北京100094

出　　处：《农业生物技术学报》2004年第2期175-178,共4页Journal of Agricultural Biotechnology

基　　金：北京市自然科学基金(No. 5022006);国家重点基础研究发展规划(973)项目(No. G1999011901)资助。

摘　　要：利用噬菌体随机7肽库分析猪繁殖与呼吸综合征病毒(PRRSV)N蛋白B细胞抗原表位。以PRRSV BJ-4株N蛋白的单克隆抗体(mAb)GE3作为筛选分子，筛选噬菌体展示的随机7肽库。经序列测定和分子生物学软件分析，表明噬菌体展示的外源氨基酸序列与PRRSV BJ-4株N蛋白的50～55aa的氨基酸序列有极高的同源性。ELISA分析结果表明，筛选到的噬菌体能特异性地与PRRSV阳性血清结合，从而证实了该表位是单克隆抗体GE3所识别的抗原表位。Phage display technology has been used successfully to identify short peptides that bind to target molecules such as monoclonal antibody (mAb), polyclonal antibody and other molecules. In order to get the epitope identified by PRRSV BJ-4 mAb GE3, a phage random heptapeptide library was biopanned with GE3. After three rounds of screening-washing-enriching, 8 phages were sequenced, and 7 clones had a consensus sequence, PEKPHF. By comparing these sequences with PRRSV nucleocapsid protein, the epitope site was located at the residue 50~55 aa of PRRSV N protein. And the results of molecular biology software analysis and ELISA suggested that the epitope was right. These results demonstrated that the sequences were the epitope of N protein of PRRSV BJ-4 isolate.

关 键 词：噬菌体随机肽库 猪繁殖与呼吸综合征病毒 PRRSV N蛋白 B细胞 抗原 表位 

分 类 号：S852.65[农业科学—基础兽医学]