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.
出处
《生物技术通报》
CAS
CSCD
2008年第6期158-163,共6页
Biotechnology Bulletin
基金
国家支撑计划项目(2006BAD06A06)
