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猪瘟病毒E2基因噬菌体展示多肽库的构建及表位鉴定
引用本文:徐和敏,王琴,徐璐,范学政.猪瘟病毒E2基因噬菌体展示多肽库的构建及表位鉴定[J].畜牧兽医学报,2010,41(1).
作者姓名:徐和敏  王琴  徐璐  范学政
作者单位:中国兽医药品监察所,猪瘟参考实验室,北京,100081
基金项目:北京市自然科学基金,"十一五"国家科技支撑计划课题,引进国际先进农业科学技术项目(2007-G57 
摘    要:本研究通过建立CSFV噬菌体展示多肽库,并对其进行生物淘选,以期获得E2蛋白上新的抗原表位.选择CSFV石门株(SM)和疫苗株(HCLV)为代表株,采用噬菌体展示技术,以T7select415-1b为载体,分别构建了CSFV SM株和HCLV株E2基因噬菌体展示多肽库(SM-E2库和HCLV-E2库).通过生物淘选和噬菌体原位杂交技术,采用7株猪瘟单克隆抗体和1株猪瘟高免血清分别对构建的SM-E2库和HCLV-E2库进行抗原表位筛选.结果共筛选到了5条与E2蛋白高度同源的序列,在E2蛋白上的同源区域分别为TAVSPTTLR、YYEP、TTWKEYSH、GGQ(V)VK和PDGLPHY.结果表明,TAVSPTTLR、YYEP和TTWKEYSH序列与目前已知E2蛋白表位一致,说明它们是E2蛋白上的优势表位;GGQ(V)VK和PDGLPHY序列与预测表位一致,推测是E2蛋白上潜在的抗原表位.

关 键 词:猪瘟病毒  E2蛋白  抗原表位  噬菌体展示多肽库

Construction of Phage Display Libraries of Classic Swine Fever Virus E2 Gene and Identification of E2 Protein Epitopes
XU He-min,WANG Qin,XU Lu,FAN Xue-zheng.Construction of Phage Display Libraries of Classic Swine Fever Virus E2 Gene and Identification of E2 Protein Epitopes[J].Acta Veterinaria et Zootechnica Sinica,2010,41(1).
Authors:XU He-min  WANG Qin  XU Lu  FAN Xue-zheng
Institution:XU He-min,WANG Qin,XU Lu,FAN Xue-zheng(National Classical Swine Fever Reference Laboratory,China Institute of Veterinary Drug Control,Beijing 100081,China)
Abstract:The phage display peptides libraries of Classic Swine fever virus(CSFV)E2 gene were constructed to identify the potential epitopes of E2 protein by biopanning.The E2 genes of Shimen strain(SM) and hog cholera lapinized virus(HCLV) strain were cloned into T7seclect-415b vector respectively.So the E2 gene phage display peptides libraries of Shimen strain and HCLV strain were constructed and named as SM-E2 libraries and HCLV-E2 libraries.CSFV polyclonal antibody(PcAB) and seven strains of McAbs were used to sc...
Keywords:classical swine fever virus  E2 protein  epitope  phage display peptides library
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