FMDV OA/58病毒株VP1蛋白结构构建与B细胞抗原表位的预测

Construction of VP1 Protein and Prediction of B Cell Epitopes in VP1 from a Foot-and-mouth Disease Virus Strain OA/58

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pGEM-T easy载体连接并转化JM109菌株,提取的重组质粒用凝胶电泳、PCR和EcoR I酶切法鉴定。运用同源模建得到OA/58 VP1蛋白3D结构,并结合理化性质、亲水性、可塑性和免疫原性进行分析,预测OA/58 VP1的抗原表位。结果OA/58 VP1存在多个潜在的抗原表位位点,可能的蛋白质抗原表位区域:2～11,15～35,38～50,77～88,90～107,121～125,131～135,140～149,154～163,169～175,178～189,197～213。应用同源模建得到的OA/58 VP1蛋白3D模型来预测其B细胞表位,为进一步研究OA/58 VP1功能,构建突变体和选择表达新型OA/58 VP1蛋白分子提供有参考价值的信息。

Foot-and-mouth disease virus strain OA/58 RNAs were used as templates for RT-PCR. The amplified cDNA products were cloned into pGEM-T easy vectors and transformed into JM109. The recombinant plasmids were identified by electrophoresis,PCR,and EcoR I cleavage. By means of homology modeling the FMDV strain OA/58 VP1 protein, the 3D mold was obtained. In order to find out B cell epitopes of OA/58 VP1, several methods were analyzed including physical and chemical characters, hydrophilicity, antigenicity. Many distinct antigenic epitopes in FMDV OA/58 VP1 were identified by computation： 2 - 11,15 - 35,38 - 50,77 - 88,90 - 107,121 - 125,131 - 135,140 - 149,154 - 163,169- 175,178- 189,197- 213. Application of homology molding OA/58 VP1 to predict B cell epitopes is reasonable. This method offers reasonable information for researching function of OA/58 VP1, constructing its variant body and selecting new expression forms of OA/58 VP1.