传染性腔上囊病病毒VP3蛋白模拟表位的筛选  

Screening for mimotopes of infectious bursal disease virus VP3

作　　者：邓小芸[1] 高玉龙[1] 高宏雷[1] 祁小乐[1] 程宇[1] 王晓艳[1] 王笑梅[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室禽传染病研究室,黑龙江哈尔滨150001

出　　处：《中国兽医科学》2006年第10期791-794,共4页Chinese Veterinary Science

基　　金：国家重点基础研究发展规划(973)项目(2005CB523202)

摘　　要：为了研究传染性腔上囊病病毒(IBDV)的致病机理及研制有效的IBDV疫苗,利用噬菌体展示技术对IBDV VP3抗原表位进行了筛选。以4株IBDV VP3单克隆抗体HRB-3F、HRB-7B、HRB-7C和HRB-10E作为筛选分子,对噬菌体随机15肽库进行3轮吸附-洗脱-扩增淘洗,从每株单克隆抗体筛选到的噬菌斑中随机挑取20个单克隆噬菌斑,通过间接ELISA和竞争抑制ELISA检测,共选出13个单克隆噬菌斑,经噬菌体gⅢ部分基因的核苷酸序列测定,确定了8个15肽为IBDV抗原表位。这8个15肽在一级结构上没有3个以上连续氨基酸与IBDV Gx(Gen-Bank登录号:AY444873)VP3的氨基酸序列相同,但二级结构上均以β折叠为主,并且与单抗的结合可被VP3蛋白有效地抑制。证实,筛选的是IBDV VP3的模拟表位。In order to study mechanism of pathogenesis and to develop an effective vaccine, the epitope of IBDV VP3 was screened by phage display. Four monoclonal antibodies to protein VP3 of infectious bursal disease virus（IBDV）,HRB-3F,HRB-TB, HRB-7C and HRB-10E, were used to screen for binding peptides from peptide 15-mer phage random peptide library. After three rounds of panning（absorption-elutionamplification） ,twenty phages for each monoclonal antibody were detected with indirect ELISA and competitive inhibition ELISA, then 13 phages were sequenced. Eight 15-mer phage peptides were determined as antigenic epitope of IBDV. Compared with sequences of IBDV Gx registered in GenBank（accession number. AY444873）, no more than 3 continuous amino acid residues were homologous with the sequences of IBDV in the primary structure, but they mainly formed beta-sheet in secondary structure and the binging to McAb can be inhibited by VP3 protein. The results indicated that the epitopes are mimotopes of IBDV.

关 键 词：传染性腔上囊病病毒 VP3蛋白 抗原表位 噬菌体随机15肽库 筛选 

分 类 号：S852.659.4[农业科学—基础兽医学]