口蹄疫病毒结构蛋白VP1上B细胞表位的筛选鉴定  

Screening and Identification of B Cell Epitopes of VP1 of Foot-and-mouth Disease Virus

作　　者：张昱[1] 王永录[1] 张永光[1] 方玉珍[1] 潘丽[1] 蒋守田[1] 吕建亮[1] 刘力宽[1] 张中旺[1] 张淑刚[1] 李正丰[1] 杜进鑫[1] 

机构地区：[1]中国农业科学院兰州兽医研究所,家畜疫病病原生物学国家重点实验室,农业部畜禽病毒学重点开放实验室,甘肃兰州730046

出　　处：《华北农学报》2009年第5期81-85,共5页Acta Agriculturae Boreali-Sinica

基　　金：国家支撑计划项目(2006BAD06A06)

摘　　要：以口蹄疫病毒株AF72 RNA为模板,反转录并扩增结构蛋白VP1基因,PCR纯化产物与pGEM-T easy载体连接并转化JM109菌株,对经凝胶电泳、PCR和EcoRⅠ酶切法鉴定为阳性的重组质粒进行测序,通过序列比对获得AF72VP1的核苷酸序列和推导氨基酸序列,综合分析结构蛋白VP1的亲水性、可塑性、抗原指数以及表面可能性等参数,预测其潜在B细胞抗原表位并人工合成表位肽段,利用间接ELISA对潜在表位肽段进行筛选鉴定,结果显示,表位VP1a和VP1d为病毒株AF72结构蛋白VP1的优势B细胞表位,该结果为进一步的FMDV多表位疫苗研究提供有价值的参考依据。Foot-and-mouth disease virus strain AF72 RNAs were used as templates for RT-PCR to amplify the VP1 gene.The purified PCR products were cloned into pGEM-T easy vectors and transformed into E.coli JM109.The positive recombinant plasmids identified by electrophoresis,PCR,and EcoRⅠcleavage were sequenced.The nucleotide and were obtained by comparing with the full-length sequence of the other reference strains.Potential B-cell epitopes of VP1 were predicted and epitope peptide segments were synthesized. They were identified by indirect ELISA. The result showed that VPla and VPld were predominant B-cell epitope of VP1, which provided valuable information for further study on the vaccine of FMD.

关 键 词：口蹄疫病毒 结构蛋白VP1 间接ELISA B细胞表位 

分 类 号：Q78[生物学—分子生物学]