口蹄疫病毒3D聚合酶B细胞表位的筛选鉴定  

Screening and Identification of B Cell Epitopes of 3D Polymerase of FMDV

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,甘肃兰州730046

出　　处：《华北农学报》2009年第3期69-73,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3D聚合酶基因,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AF723D聚合酶的核苷酸序列和推导氨基酸序列,综合分析3D聚合酶的亲水性、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对潜在表位进行筛选鉴定,结果显示,表位3D2和3D4为病毒株AF72 3D聚合酶的优势B细胞表位,该结果将为进一步的FMDV多表位疫苗研究奠定基础。Foot-and-mouth disease virus strain AF72 RNAs were used as templates for RT-PCR to amplify the 3D gene.The purified PCR products were cloned into pGEM-T easy veetors and transformed into E. coli JM109.The positive recombinant plasmids identified by electrophoresis, PCR, and Eco R Ⅰ cleavage were sequenced. The nucleotide and amino acid sequence were obtained by comparing with the full-length sequence of the other reference strains. Potential B cell epitopes of 3D were predicted and epitope peptide segments were synthesized. They were identified by indirect ELISA. The result showed that 3D2 and 3D4 were predominant B cell epitope of 3D,it provided a basis for further study on the multi-epitope vaccine of FMD.

关 键 词：口蹄疫病毒 3D聚合酶 间接ELISA B细胞表位 

分 类 号：Q78[生物学—分子生物学]