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目的获得EV71P1保守氨基酸序列COBRA EV71 P1和基于此序列的B细胞抗原表位预测信息。方法从NCBI(http://www.ncbi.nlm.nih.gov/)上下载获取1970-2010年间世界各地暴发流行的包含P1或VP1基因片段的EV71源序列和对应的氨基酸序列,根据VP1基因片段信息构建系统进化树,对源序列进行基因分组;对多基因型的EV71 P1氨基酸源序列采用多序列比对,以优势氨基酸位点代替差异氨基酸位点的方法,获得EV71 P1保守序列(COBRA EV71 P1),并运用IEDB Analysis Resource (http://tools.immuneepitope.org/main/)在线预测工具和DNASTAR软件里的Protean模块,运用单参数和二级结构预测综合考虑的方法,对COBRA EV71 P1前体蛋白的B细胞抗原表位进行预测。结果成功比对出COBRA EV71 P1氨基酸序列,预测结果发现在COBRA EV71 P1前体蛋白的第10-24,41-60,68-82,208-225,328-345,498-514,592-609,664-678,772-786和841-855位点均具有较好的亲水性、表面可及性、柔韧性和抗原指数,并且在二级结构上含有易形成抗原表位的无规则卷曲和β-转角,有可能是COBRA EV71 P1前体蛋白的优势表位。结论经生物信息学分析,推测COBRA EV71 P1可能具有三个基因型EV7 1P1前体蛋白的候选B细胞线性抗原表位,为EV71重组多表位疫苗设计和多基因型病毒样颗粒疫苗实验提供了理论基础。
OBJECTIVE: To obtain COBRA EV71 P1, a conserved amino acid sequence of EV71P1, and B cell epitope predictions based on this sequence. Methods The EV71 source and corresponding amino acid sequences of the EV71 gene fragment containing the P1 or VP1 gene outbreaks worldwide from 1970 to 2010 were downloaded from NCBI (http://www.ncbi.nlm.nih.gov/). According to the VP1 gene The phylogenetic tree was constructed based on the phylogenetic tree and the gene sequences of the EV71 P1 gene were grouped. Multiple sequence alignment of the EV71 P1 amino acid sequence of multiple genotypes was performed. The conservative amino acid sequence of EV71 P1 was substituted for the amino acid sequence of EV71 P1 (COBRA EV71 P1) and using the IEDB Analysis Resource (http://tools.immuneepitope.org/main/) online prediction tool and the Protean module in the DNASTAR software, using the single-parameter and secondary-structure prediction methods to consider COBRA EV71 P1 precursor protein B cell epitopes predicted. Results The amino acid sequence of COBRA EV71 P1 was successfully aligned. The predicted results were found to be 10-24, 41-60, 68-82, 208-225, 328-345, 498-514, 592-609, 664-678, 772-786 and 841-855 of the COBRA EV71 P1 precursor protein Sites have good hydrophilicity, surface accessibility, flexibility and antigen index, and in the secondary structure contains easy to form random coil epitopes and β-turn, it may be COBRA EV71 P1 before The dominant epitope of the body protein. Conclusions Bioinformatics analysis suggested that COBRA EV71 P1 may have candidate B cell linear epitopes of three genotype EV7 1P1 precursor proteins, which provided the experimental basis for EV71 recombinant multi-epitope vaccine design and multi-genotype virus-like particle vaccine experiment Theoretical basis.