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.
出处
《华北农学报》
CSCD
北大核心
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)
