溶藻弧菌Ⅲ型分泌系统效应蛋白Va1686基因的克隆和生物信息学分析  

Cloning and Bioinformatics Analysis of Effector Protein Va1686 Gene in Type Ⅲ Secretion System of Vibrio alginolyticus

作　　者：宋大伟[1,2,3] 庞欢瑛 邱明生[1,2,3] 林紫薇 汤菊芬[1,2,3] 梁珊珊 简纪常[1,2,3] Song Dawei 1,2,3 ,Pang Huanying 1,2,3, Qiu Mingsheng 1,2,3 ,Lin Ziwei 1,2,3, Cai Shuanghu 1,2,3 ,Cai Jia1,2,3 ,Tang Jufen1,2,3, Jian Jichang1,2,3(1 Fisheries College of Guangdong Ocean University, Zhanjiang, 524088; 2 Guangdong Provincial Key Laboratory of Pathogenic Biology and Epidemilogy for Aquatic Economic Animals, Zhanjiang, 524088; 3 Key Laboratory of Control for Diseases of Aquatic Economic Animals of Guangdong Higher Education Institutes, Zhanjiang, 52408)

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3期1210-1217,共8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);广东省省级科技计划项目(2014A020208117)共同资助

摘　　要：本研究从溶藻弧菌中成功克隆获得Va1686基因的编码区序列（GenBank：KX245316），该基因全长1164bp，可编码387个氨基酸，应用生物信息学的方法和工具对溶藻弧菌HY9901III型分泌系统效应蛋白Va1686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：Va1686蛋白为稳定的亲水性蛋白，蛋白呈酸性，不具有跨膜区和信号肽，二级结构以仅螺旋为主。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。Va1686蛋白包含一段与细胞分裂相关的Fic superfamily保守结构域。生物信息学分析表明Va1686可能的B细胞抗原优势表位，分别是第48～49、82～85、125～126、150～153、185～186、236～237等区段。利用SWISS-MODEL软件，模拟了Va1686亚基三维结构模型，发现与其同源蛋白副溶血弧菌vopS非常相似，相似度达到89．46％。本研究从生物信息学角度分析了Va1686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： KX245316） of Va1686 gene was cloned from Vibrio alginolyticus. The total length of the gene was 1 164 bp, and it could encode 387 amino acids. The physicochemical properties, protein structure, genetic evolutionary relationship and antigenic characteristics of the effector protein Va1686 of HY9901 type Ⅲ secretion system in Vibrio alginolytieus were studied and analyzed by bioinformatics methods and tools. The results showed that Va1686 was a stable hydrophilic and acidic protein without a transmembrane region and a signal peptide, and secondary structure was rich in α-helix. The evolutionary analysis showed that Vibrio alginolyticus HY9901 and V. harveyi were clustered together, which indicated that the genetic relationship between the two species was closest. Va1686 contained a Fic superfamily conserved domain associated with cell division. Bioinformatics analysis showed that the B-cell preponderant epitopes of Va1686 might be located in the regions of 48-49, 82-85, 125-126, 150-153, 185-186, 236-237 and so on. The 3D structure model of Va1686 subunit was simulated by SWISS-MODEL software and it was found that Va1686 and the homologous protein vopS were similar and the similarity was 89.46%. In this study, the feasibility of Va1686 as a common antigen of Vibrio was analyzed from the perspective of bioinformatics, which might lay the foundation for the next step in vaccine development.

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：S941.4[农业科学—水产养殖]