溶藻弧菌Ⅲ型分泌系统效应蛋白HY322基因的克隆和生物信息学分析  

Cloning and Bioinformatics Analysis of Type Ⅲ Secretion System Effector Protein HY322 Gene of Vibrio alginolyticus

作　　者：宋大伟[1,2,3] 庞欢瑛 邱明生[1,2,3] 林紫薇[1,2,3] 蔡双虎 蔡佳[1,2,3] 汤菊芬 简纪常[1,2,3] Song Dawei;Pang Huanying;Qiu Mingsheng;Lin Ziwei;Cai Shuanghu;Cai Jia;Tang Jufen;Jian Jichang(Fisheries College of Guangdong Ocean University, Zhanjiang, 524088;Guangdong Provincial Key Laboratory of Pathogenic Biology and Epidemilogy for Aquatic Economic Animals, Zhanjiang, 524088;Key Laboratory of Control for Diseases of Aquatic Economic Animals of Guangdong Higher Education Institutes, Zhanjiang, 524088)

机构地区：[1]广东海洋大学水产学院,湛江524088 [2]广东省水产经济动物病原生物学及流行病学重点实验室,湛江524088 [3]广东省水产经济动物病害控制重点实验室,湛江524088

出　　处：《基因组学与应用生物学》2018年第7期2843-2849,共7页Genomics and Applied Biology

基　　金：国家自然科学基金(31402344;31572656);广东省自然科学基金重大培育项目(2015A030308020)共同资助

摘　　要：本研究从溶藻弧菌中成功克隆获得HY322基因的编码区序列（GenBank：KX245317.1）,该基因全长969 bp,可编码322个氨基酸,应用生物信息学的方法和工具对溶藻弧菌HY9901效应蛋白HY322的理化性质、蛋白结构、遗传进化关系和抗原特性等方面进行预测和分析。结果表明：HY322蛋白为不稳定的亲水性蛋白,蛋白呈酸性,无跨膜区,无明显的信号肽;二级结构以α螺旋为主,进化分析显示溶藻弧菌HY9901与哈维氏弧菌（V.harveyi）聚为一支,表明在进化关系上最为接近。HY322序列中包含有一个与鞭毛形成有关的Fli N功能结构域。多种参数预测了HY322可能的B细胞抗原优势表位,分别是第32~33、100~102、138~140、215~216、235~238、246~249区段。利用SWISS-MODEL软件,得到了HY322亚基三维模型。本研究从生物信息学角度验证了HY322作为弧菌共同抗原的可行性,为下一步的疫苗研发提供了理论依据。In this study, the coding sequence （GenBank： KX245317.1） ofHY322 gene was cloned from Vibrio alginolyticus. The total length of its gene was 969 bp, encoding 322 amino acids. The physicochemical properties, protein structure, genetic evolutionary relationship and antigenic characteristics of the effector protein HY322 of vibrio alginolyticus HY9901 were predicted and analyzed by bioinformatics methods and tools. The results showed that HY322 protein might be an unstable hydrophilic and acidic protein without a transmembrane region or a signal peptide, and the secondary structure was mainly α-helix. The evolutionary analysis showed that Vibrio alginolyticus HY9901 and V. harveyi were clustered together, which indicated that their genetic relationship was the closest. The HY322 sequence contained a FliN superfamily conserved domain associated with Flagellar motor switch. Various parameters predicted that the B-cell preponderant epitopes of HY322 might be localized in the regions of 32-33, 100-102, 138-140, 215-216, 235-238, 2464249. The 3D structure model of HY322 subunit was simulated by SWISS-MODEL software. This study verified the feasibility of HY322 as a common antigen of Vibrio from the perspective of bioinformatics, which might provide a theoretical basis for the further research and development of vaccine.

关 键 词：溶藻弧菌 三型分泌系统 效应蛋白 生物信息学分析 

分 类 号：S941.42[农业科学—水产养殖]