CAJ | 학술논문

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AF72 VP1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.
이구제역병독주AF72 RNA위모판,반전록병확증결구단백VP1기인,PCR순화산물여pGEM-T easy재체련접병전화JM109균주,대경응효전영、PCR화EcoRⅠ매절법감정위양성적중조질립진행측서,통과서렬비대획득AF72 VP1적핵감산서렬화추도안기산서렬,종합분석결구단백VP1적친수성、가소성、항원지수이급표면가능성등삼수,예측기잠재B세포항원표위병인공합성표위태단,이용간접ELISA대잠재표위태단진행사선감정,결과현시,표위VP1a화VP1d위병독주AF72결구단백VP1적우세B세포표위,해결과위진일보적FMDV다표위역묘연구제공유개치적삼고의거.
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 Eco R I cleavage 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.