噬菌体展示随机十肽库的构建及抗WSSV单链抗体-VirologicaSinica.PDF

第 21 卷 6 期 中 国 病 毒 学 21 （6 ）：614-618 2006 年 11 月 VIROLOGICA SINICA November 2006 噬菌体展示随机十肽库的构建及抗 WSSV 单链抗体A1 的模拟抗原表位的淘选和鉴定* 1,2 1,2 1,2 1 1** 袁 丽 ，王玉珍 ，肖 囡 ，张晓华 ，戴和平 （1.中国科学院水生生物研究所，湖北武汉 430072；2.中国科学院研究生院，北京 100039） Construction of a Phage Display Random Decapeptide Library and Panning for Mimic Antigen Epitope of ScFv A1 Against WSSV 1, 2 1, 2 1, 2 1 1** YUAN Li ，WANG Yu-zhen ，XIAO Nan ，ZHANG Xiao-hua , DAI He-ping (1.Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China; 2. Graduate School of the Chinese Academy of Sciences ，Beijing 100039, China ) Abstract ：A phage display random decapeptide library with high titer was constructed using pCANTAB 5 E as vector and used to select and identify antigen epitopes. ScFv A1, which specifically binds the shrimp’s White spot syndrome virus (WSSV), was used to pan against the decapeptide library and a random 15mer library, respectively. A series of positive clones that bind specifically to scFv A1 were obtained. The amino acid sequences of the positive peptides were compared with WSSV388 fragment, which is the antigen of scFv A1. We found that most of the sequences of the positive peptides were similar to a fragment ，K••••R••R•Q, located on C-terminal of WSSV388 fragment. We deduced that the mimic antigen epitopes of scFv A1 were conformational epitopes structured by the discontinuous amino acid fragment, but not by a line