应用CSFV-E2基因噬菌体肽库筛选猪瘟病毒抗原表位  

Screening of epitopes of E2 protein using phage display libraries of E2 gene of classic swine fever virus

作　　者：汤波[1,2] 范学政[2] 徐璐[2] 王琴[2] 黄伟[1] 刘俊[1,2] 沙莎[1] 陈蕾[2] 周远成[2] 

机构地区：[1]西南大学动物科技学院,重庆荣昌402460 [2]中国兽医药品监察所国家猪瘟参考实验室,北京海淀100081

出　　处：《中国兽医杂志》2009年第4期12-15,共4页Chinese Journal of Veterinary Medicine

基　　金：北京市自然科学基金(5072041);“十一五”国家支撑计划课题(2006BAD06A03)

摘　　要：采用猪瘟单抗对猪瘟病毒石门株E2基因噬菌体随机肽库(SM-E2库)进行淘洗以研究猪瘟病毒E2抗原表位。运用猪瘟单克隆抗体WH303、WH211和M56对SM-E2库进行4轮生物淘洗,对阳性克隆再经噬菌体原位杂交、特异PCR测序分析,然后人工合成阳性多肽进行ELISA反应。经鉴定分析发现单抗WH303从SM-E2库中筛选得到一段CSFV特异的抗原多肽IECTAVSPTTLRTEVVKT,并且该段多肽与已知CSFV表位TAVSPTTLR极为相似;单抗WH211和M56未能筛选到包含CSFV序列的克隆。结果表明,利用靶基因噬菌体随机多肽库筛选抗原表位能够得到更接近于真实表位的抗原表位。E2 gene phage display peptides libraries of CSFV Shimen strain （SM-E2 libraries） were selected by CSFV monoclonal antibodies （MAbs） to screen the potential epitopes of CSFV-E2 protein. Four rounds of biopannings were carried out to the SM-E2 libraries using MAbs WH303, WH211, M56. The positive clones were identified by phage in Situ Hybridization, Sequence analysis and ELISA. After analyzing, the one sequence which was highly homologous with TAVSPTTLR, were screened by the MAbs 303. None was gained by WH211 and M56. Results suggested that the target gene phage-display library of random peptides is a novel tool for investigation into Antigenic epitopes.

关 键 词：猪瘟病毒 抗原表位 噬菌体随机肽库 

分 类 号：S852.651[农业科学—基础兽医学]