鹅细小病毒VP3蛋白B细胞线性表位的精确定位  

Exact Location of Linear B-cell Epitopes of VP3 Protein of Goose Parvovirus

作　　者：郭鹭[1] 鞠环宇[1] 于天飞[1] 荆志强[1] 马波[1] 王君伟[1] 

机构地区：[1]东北农业大学动物医学学院,哈尔滨150030

出　　处：《畜牧兽医学报》2010年第8期988-994,共7页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：黑龙江省教育厅重大项目(1054lz004);黑龙江省攻关项目(GB01B503-02GB04B504)

摘　　要：本试验旨在利用获得的4株抗鹅细小病毒(GPV)VP3的单克隆抗体,进一步对GPV VP3 B细胞线性抗原表位精确定位。根据笔者实验室已鉴定的线性抗原表位区结果,筛选出单抗所针对的优势抗原表位区。设计并合成互相重叠5 aa的10 aa短肽寡聚核酸片段,退火后,连入pET-32a载体,经转化鉴定,诱导表达后,获得相应的小片段融合蛋白,并利用单抗通过Western blot进行抗原性鉴定。同样方法进行短肽片段两端氨基酸的逐个缺失设计,进一步精确定位,结果鉴定出2个抗原表位,分别为430-435 aa和643-647 aa。Four monoclonal antibodies against Goose Parvovirus（GPV） VP3 already obtained by our laboratory were used for the exact location to linear B-cell epitopes against VP3 of GPV.On the basis of the epitopes results of GPV VP3 protein that have already been identified in previous study,screened epitopes which can be identified by monoclonal antibodies directly.Oligomeric nucleic acid fragments of ten amino acid fragments mutually coinciding five amino acids were contrived,after annealing,connected with pET-32a,by allaxis identification,derivation expressed,small fragment fusion proteins were obtained.The antigenicity was identified by Western blot with McAbs.With the same method,amino acids were deleted one by one,and two epitopes were located at 430-435 aa and 643-647 aa.

关 键 词：鹅细小病毒 VP3蛋白 单克隆抗体 B细胞线性表位 

分 类 号：S852.659.2[农业科学—基础兽医学]