文献类型：期刊文献

中文题名：猪瘟病毒E2蛋白配体表位的筛选与鉴定

英文题名：Identification of the ligand epitope of E2 protein on classical swine fever virus

作者：银梅[1];李鹏[2];岳锋[2];张秋雨[1];胡东方[1];宁红梅[1];孔令芸[3];姜金庆[1];王选年[2]

第一作者：银梅

机构：[1]河南科技学院动物科技学院,河南新乡453003;[2]新乡学院生命科学技术学院,河南新乡453003;[3]新乡县农牧局(新乡县林业局),河南新乡453700

第一机构：河南科技学院动物科技学院,河南新乡453003

年份：2019

卷号：0

期号：6

起止页码：1045-1051

中文期刊名：中国兽医学报

外文期刊名：Chinese Journal of Veterinary Science

收录：CSTPCD;;北大核心:【北大核心2017】；CSCD:【CSCD_E2019_2020】；

基金：国家自然科学基金资助项目(31201877);河南省教育厅创新人才资助项目(14HASTIT026)

语种：中文

中文关键词：猪瘟病毒;E2蛋白;配体表位

外文关键词：classical swine fever virus;E2 protein;ligand epitope

摘要：为进一步精确定位猪瘟病毒(CSFV)E2蛋白配体表位信息,利用DNASIS(Ver.2.5)软件分析设计合成7条多肽,主要覆盖E2蛋白,分别命名为SE21、SE22、SE231、SE232、SE241、SE242、SE03,同时设计合成1条无关多肽CP,均标记FITC。将PK-15细胞作为靶细胞,通过多肽与PK-15细胞结合试验、多肽抑制CSFV感染PK-15细胞试验和CSFV阻断多肽与PK-15细胞结合试验,筛选和鉴定E2蛋白配体表位。结果表明,多肽SE241、SE242与PK-15细胞结合的平均荧光强度分别为74.43±4.59和83.60±3.11,显著高于其他多肽,抑制CSFV感染PK-15细胞的感染抑制率也显著高于其他多肽,同时这2条多肽与PK-15细胞的结合被CSFV有效阻断。证实SE241、SE242多肽为CSFV与靶细胞PK-15细胞结合的配体表位,并能抑制CSFV感染PK-15细胞。本试验为CSFV新型多肽疫苗或免疫佐剂的研制提供依据。
In order to further accurately ascertain the E2 protein ligand epitope information,seven polypeptides were designed and synthesized by DNASIS(Ver.2.5)software.They mainly covered with E2 protein of classical swine virus(CSFV),named SE21,SE22,SE231,SE232,SE241,SE242,SE03.At the same time,an unrelated polypeptide CP was designed and synthesized,and the all were labeled FITC,PK-15 cells were used as target cells.And the E2 protein ligand sites were screened and identified through the PK-15 cell binding test,inhibition of CSFV infection by peptides and binding assay of CSFV blocking peptide with PK-15 cells.Results indicated that the average fluorescence intensity of the peptide SE241 and SE242 combined with PK-15 cells were 74.43±4.59,and 83.60±3.11,which were significantly higher than that of other polypeptides.The inhibitory rate of CSFV infection on PK-15 cells was also significantly higher than that of other polypeptides and the binding of these two polypeptides to PK-15 cells was effectively blocked by CSFV.It was confirmed that SE241 and SE242 polypeptides were ligand epitopes of CSFV binding to target cells PK-15 cells,and could inhibit CSFV infection of PK-15 cells.This study further reveals the molecular basis of CSFV infection in target cells and provides theoretical support for the development of novel peptide vaccines or immune adjuvants for CSFV.