中蜂囊状幼虫病毒LN-QY株VP1蛋白的空间结构与B细胞抗原表位预测

Predication of Spatial Structure and B Cell Epitope of VP1 Protein of Chinese Sacbrood Virus LN-QY Strain

目的预测中蜂囊状幼虫病毒(Chinese sacbrood virus,CSBV)LN-QY株VP1蛋白的空间结构及其B细胞抗原表位。方法以CSBV LN-QY株RNA为模板,RT-PCR扩增结构蛋白VP1基因,与pMD18-T载体连接后,转化感受态大肠杆菌DH5α,对经EcoRⅠ和HindⅢ双酶切鉴定为阳性的重组质粒进行测序,通过序列比对,获得LN-QY株VP1的核苷酸序列和推导的氨基酸序列,建立VP1结构蛋白的3D模型,在此基础上,应用DNAStar软件中的Protean模块综合分析结构蛋白的柔性区域,亲水性,表面可能性以及抗原指数等参数。结果 VP1结构蛋白的空间构象比较规则,其可能的B细胞抗原表位区域位于47-53,139-145,273-284氨基酸区段。结论本研究为CSBV LN-QY株VP1蛋白表位疫苗的设计以及血清诊断试剂盒的研制奠定了理论基础。

Objective To predict the spatial structure and B cell epitope of VP1 protein of Chinese sacbrood virus（CSBV） LN-QY strain.Methods The gene encoding VP1 protein of CSBV was amplified by RT-PCR using the RNA of LN-QY strain as a template,and inserted into vector pMD18-T.The constructed recombinant plasmid was transformed to competent E.coli DH5α,and the positive clones identified by digestion with EcoRⅠ and Hind Ⅲ was sequenced.The nucleotide and deduced amino acid sequences were obtained by comparing the sequences with those of other reference strains,and the 3D model of structural protein of VP1 was established,based on which several parameters of the structural protein,such as flexible region,hydrophilicity,surface probability and antigenic index were analyzed by the Protean module in DNAStar software.Results The spatial confirmation of VP1 protein was relatively regular,in which the potential B cell antigenic epitopes were located at 47 ～ 53,139 ～ 145 and 273 ～ 284 aa.Conclusion The study provided a theoretical basis for design of VP1 epitope vaccine against CSBV LN-QY strain as well as the preparation of serological diagnostic kit.