口蹄疫病毒株AF72 VP1的结构构建与B细胞表位预测  

Construction of VP1 and Prediction of B Cell Epitopes from FMDV AF72

作　　者：张昱[1] 王永录[1] 张永光[1] 潘丽[1] 方玉珍[1] 刘力宽[1] 蒋守田[1] 吕建亮[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,兰州730046

出　　处：《生物技术通报》2008年第6期158-163,共6页Biotechnology Bulletin

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,反转录并扩增目的基因,PCR纯化产物与pGEM-T easy载体连接并转化JM109菌株,用凝胶电泳、PCR和EcoRⅠ酶切法鉴定为阳性的重组质粒进行测序。比对测序结果确定AF72 VP1的核苷酸序列,利用同源建模的方法建立AF72 VP1结构蛋白的3D结构,在此基础上,综合亲水性、可塑性、抗原指数以及表面可能性等参数预测AF72 VP1结构蛋白的B细胞抗原表位。结果显示,VP1结构蛋白呈现较规则的空间构象,其中5-11、24-30、43-47、82-87、132-147和196-203氨基酸区段是AF72 VP1结构蛋白可能的B细胞抗原表位区域,该结果将为进一步的FMDV多表位疫苗研究提供很有价值的参考信息。Foot-and-mouth disease virus strain AF72 RNAs were used as templates for RT-PCR to amplify the target gene.The purified PCR products were cloned into pGEM-T easy Vectors and transformed into E.coli JM109.The positive recombinant plasmids which have been identified by electrophoresis,PCR and EcoRⅠcleavage respectively,were sequenced.The nucleotide sequence were confirmed by comparing with the full-length sequence of the other reference strains.By homology modeling,the 3D model of AF72 VP1 structure protein was obtained.Then several parameters,including hydrophilicity,flexibility,antigenic index and surface probability were integrated,and B-cell epitopes in VP1 were predicted.The result showed the conformation of AF72 VP1 was regular,and B-cell epitopes in VP1 probably existed in the following regions,including 5aa-11aa,24aa-30aa,43aa-47aa,82aa-87aa,132aa-147aa and 196aa-203aa.This result offers valuable information for further research of FMDV multi-epitope vaccine.

关 键 词：口蹄疫病毒 VP1结构蛋白 3D结构 B细胞表位 

分 类 号：S852.5[农业科学—基础兽医学] Q789[农业科学—兽医学]