非洲猪瘟病毒p72蛋白抗原表位预测分析及多表位疫苗的构建

Epitope Prediction Analysis of African Swine Fever Virus p72 Protein and Construction of Multi-epitope Vaccine

为了预测非洲猪瘟病毒2018/AnhuiXCGQ株p72蛋白的T、B淋巴细胞抗原表位,并完成多表位疫苗的构建。试验采用生物信息学技术ExPASy、SOMPA、PSIPRED Server、DNASTAR及Phyre对该蛋白的理化性质、二级结构、三级结构进行初步预测分析,运用ABCpred Prediction、Scratch、NetCTL及IEDB预测其T、B淋巴细胞表位,最终设计构建得到多表位疫苗。依据各生物学方法分析预测,得到B淋巴细胞优势表位:110~120、77~88、45~55、12~18、27~37位置;T淋巴细胞优势表位298~307、520~531、203~212位置,并设计得到T、B淋巴细胞表位表达盒,ExPASy预测表示该表位盒为亲水性蛋白。以此多表位表达盒为目的基因,以pET-28a构建设计多表位疫苗。表明该蛋白有多个潜在抗原表位,并可依据预测得到的蛋白二级结构、三级结构等参数信息及多个预测抗原位点构建ASFV多表位疫苗,表达纯化用于免疫试验。

To predict the T and B lymphocyte epitopes of p72 protein of African swine fever virus 2018/AnhuiXCGQ strain,and complete the construction of multi-epitope vaccine.The bioinformatics technologies ExPASy,SOMPA,PSIPRED Server,DNAstar and Phyre were used to predict the physicochemical properties,secondary structure and tertiary structure of the protein.The predictions were based on ABCpred Prediction,Scratch,NetCTL and IEDB.The T and B lymphocyte epitopes were finally designed and constructed to obtain a multi-epitope vaccine.According to the analysis and prediction of various biological methods,the dominant epitopes of B lymphocytes were obtained:110~120,77~88,45~55,12~18,27~37;T lymphocyte dominant epitopes were 298~307,520~531,and 203~212.T and B lymphocyte epitope expression cassettes were designed,and ExPASy prediction indicated that the epitope cassette was a hydrophilic protein.Using this multi-epitope expression cassette as the target gene,a multi-epitope vaccine was designed using pET-28 a.This study indicated that the protein has multiple potential epitopes,and can be used to construct ASFV multi-epitope vaccine based on predicted protein secondary structure,tertiary structure,other parameter information,multiple predicted antigenic sites and immunization test.