首页|嗜水气单胞菌外膜蛋白TolB的原核表达及生物信息学分析

嗜水气单胞菌外膜蛋白TolB的原核表达及生物信息学分析

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为筛选TolB作为嗜水气单胞菌(Aeromonas hydrophila)渔用疫苗候选抗原,从嗜水气单胞菌中克隆了外膜蛋白tolB基因并异源表达,采用生物信息学方法预测TolB的理化性质和结构特征.结果显示,tolB基因片段(去除信号肽)全长1263 bp,编码1条含有420个氨基酸残基的多肽,分子质量45.78 ku.成功在大肠杆菌E.coli BL21(DE3)中异源表达TolB蛋白,其为稳定的亲水性外膜蛋白.TolB蛋白家族在不同菌株间具有同源性,尤其在气单胞菌间亲缘关系更近.TolB蛋白二级结构以无规则卷曲为主,具有潜在的B细胞、细胞毒性T淋巴细胞(CTL)和辅助T细胞(Th)抗原表位;相互作用网络主要为Tol-Pal系统蛋白.综上,TolB具有成为嗜水气单胞菌亚单位疫苗有效候选抗原的特性.
Expression and Bioinformatics Analysis of Aeromonas hydrophila Outer Membrane TolB Protein
The purpose of this study is to evaluate the application of TolB as the candidate vaccine antigen to prevent Aeromonas hydrophila infection in fish.Outer membrane protein gene tolB from A.hydrophila was cloned and expressed in Escherichia coli BL21(DE3).Physicochemical properties and advanced structures of TolB protein were analyzed by bioinformatics.The results indicated that tolB gene had an open reading frame of 1263 bp(exclude signal peptide),encoding a protein(TolB)of 420 amino acids with molecular mass of 45.78 ku.TolB was successfully expressed and purified from Escherichia coli BL21(DE3).TolB was a stable and hydrophilic outer membrane protein.TolB had homology among different bacteria,and was more closely related in Aeromonas.The secondary structure of TolB was dominated by random coil.TolB had potential B,CTL and Th cell antigen epitopes and interacted with other Tol-Pal system proteins.In conclusion,TolB could be used as an effective vaccine candidate antigen agaisnt A.hydrophila.

Aeromonas hydrophilaTolBProkaryotic expressionProtein purificationBioinformatics

陈甜梦、蔡彤璇、王嘉璐、田牧野、赵宝华、刘东

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河北师范大学 生命科学学院,河北 石家庄 050024

嗜水气单胞菌 TolB 原核表达 蛋白质纯化 生物信息学

河北省自然科学基金项目河北省高等学校科学技术研究项目河北省教育厅优秀青年基金项目河北师范大学重点基金项目

C2019205044ZD2018070YQ2014026L2016Z03

2024

河南农业科学
河南省农业科学院

河南农业科学

CSTPCD北大核心
影响因子:0.787
ISSN:1004-3268
年,卷(期):2024.53(2)
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