细粒棘球绦虫多价EgA31-EgG1Y162抗原序列优化分析  

Optimization analysis of multivalent EgA31-EgG1Y162 antigen sequence of Echinococcus granulosus

作　　者：张杰[1] 李玉娇[2] 张峰波[1] 孔慧芳 周晓涛[1] 王红英[1] 丁剑冰[1] ZHANG Jie;LI Yu-Jiao;ZHANG Feng-Bo;KONG Hui-Fang;ZHOU Xiao-Tao;WANG Hang-Ying;DING Jian-Bing(College of Basic Medicine,Xinjiang Medical University,Urumqi 830011,China)

机构地区：[1]新疆医科大学基础医学院,乌鲁木齐830011 [2]新疆医科大学第一附属医院,乌鲁木齐830011

出　　处：《中国免疫学杂志》2020年第1期19-25,共7页Chinese Journal of Immunology

基　　金：国家自然科学基金(31560262,81660343);新疆十三五重点学科(高原)资助项目

摘　　要：目的:将优化的EgA31-EgG1Y162抗原基因构建于pET30a上,诱导蛋白表达,SDS-PAGE鉴定蛋白表达效果,用生物信息学软件分析优化的EgA31-EgG1Y162抗原序列,为构建高效的疫苗奠定理论基础。方法:运用ProtParam,DNAstar,E.coli Codon Usage Analyzer等多种生物信息学软件分析优化后EgA31-EgG1Y162蛋白的理化性质,比较其优化前后序列的异同点及抗原表位分布情况等。结果:EgA31-EgG1Y162蛋白属于不稳定性蛋白且亲水性蛋白,通过网上在线软件分析,优化序列的密码子中不含利用率低的密码子。表位预测分析确定6个T细胞表位的氨基酸区域1~15、86~97、177~193、359~388、305~319、344~358和7个B细胞表位为102~112、172~204、318~334、343~353、394~409、415~431、443~458。结论:多价EgA31-EgG1y162融合抗原蛋白的优化序列中不含利用率低的密码子,且T和B细胞表位未受影响,为重组蛋白体外的有效表达奠定实验基础,对包虫病的多价表位疫苗研究具有重要的指导作用。Objective:To construct the optimized EgA31-EgG1Y162 antigen gene on pET30a,inducing protein expression,identifing protein expression by SDS-PAGE and analyzing the optimized EgA31-EgG1Y162 antigen sequence by bioinformatics software to lay a theoretical foundation for constructing a highly efficient vaccine.Methods:The physicochemical properties of the optimized EgA31-EgG1Y162 protein were analyzed by ProtParam,DNAstar,E.coli Codon Usage Analyzer and other bioinformatics software.The similarities and differences of the Pre-optimized and optimized sequence and the distribution of antigenic epitopes were compared.Results:EgA31-EgG1Y162 protein was an unstable protein and hydrophilic protein.The optimized sequence does not contain low-coefficient codons by online software analysis.Epitope prediction analysis determined that the amino acid positions of the six T cell epitopes were 1-15,86-97,177-191,359-378,305-319,344-358 and seven B cell epitope sites were 102-112,172-204,318-334,343-353,394-409,415-431,443-458.Conclusion:The optimized sequence of the multivalent EgA31-EgG1y162 fusion antigen protein does not contain low-coefficient codons.and T and B cell epitopes are unaffected,which lays an experimental foundation for the efficient expression of recombinant proteins in vitro and plays an important guiding role in the multivalent echinococcosis epitope vaccine research.

关 键 词：细粒棘球绦虫 EgA31蛋白 EgG1Y162蛋白 表位 

分 类 号：R392.9[医药卫生—免疫学]