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。
张昱
,
王永录
,
张永光
,
方玉珍
,
潘丽
,
蒋守田
,
吕建亮
,
刘力宽
,
张中旺
,
张淑刚
,
李正丰
,
杜进鑫
. 口蹄疫病毒结构蛋白VP1上B细胞表位的筛选鉴定[J]. 华北农学报, 2009
, 24(5)
: 81
-85
.
DOI: 10.7668/hbnxb.2009.05.018
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 EcoR ivcleavage were sequenced.The nucleotide and were obtained by comparing with the ful-llength sequence of the other reference strains.Potential B-cell epitopes of VP1 were predicted and epitope peptide segments were synthesized.They were ident ified by indirect ELISA.The result showed that VP1a and VP1d were predominant B-cell epitope of VP1,which provided valuable information for further study on the vaccine of FMD.
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