口蹄疫OA／58病毒株VP2蛋白结构模拟与B细胞抗原表位的分析

以口蹄疫病毒株OA/58 RNA为模板,反转录并扩增目的cDNA,然后与pMD18-T载体连接并转化JM109菌株,提取的重组质粒用凝胶电泳、PCR和BamH I,Hind III双酶切法鉴定。运用同源模建得到OA/58 VP2蛋白三维空间结构,并结合理化性质、亲水性、可塑性和免疫原性进行分析,找出OA/58 VP2蛋白的B细胞抗原表位。结果表明,OA/58 VP2蛋白存在多个潜在的抗原表位,可能的蛋白质抗原表位区域:1~23,40~63,71~78,82~91,102~106,113~119,131~138,148~154,166~177,189~196,212~218。应用同源模建得到的OA/58 VP2蛋白三维空间结构来预测其B细胞表位,为进一步研究OA/58 VP2蛋白在引起易感宿主体液免疫应答方面提供了一种可视化的技术平台,并且为选择表达其他口蹄疫病毒株的VP2蛋白分子提供有参考价值的信息。

Construction of VP2 Protein and Prediction of B Cell Epitopes in VP2 from A Foot-and-mouth Disease Virus Strain OA/58

Foot-and-mouth disease virus strain OA/58 RNAs were used as templates for RT-PCR.The amplified cDNA products were cloned into pMD18-T vectors and transformed into JM109.The recombinant plasmids were identified by electrophoresis,PCR,and analysis of tow cleavages with BamH I and Hind III.Depending on homology modeling the FMDV strain OA/58 VP2 protein,the 3D mold was gained.To come out B cell epitopes concerning FMDV OA/58 VP2 protein,several standards were analyzed including physical and chemical characters,hydrophilicity,antigenicity.Many distinct antigenic epitopes in FMDV OA/58 VP2 protein were identified by computation:1-23,40-63,71-78,82-91,102-106,113-119,131-138,148-154,166-177,189-196,212-218.Application of homology molding OA/58 VP2 protein to predict B cell epitopes is reasonable.This method offers reasonable information for researching function of OA/58 VP2 protein,constructing its variant body and selecting new expression forms of OA/58 VP2 protein.