鳗鲡病原菌二联外膜蛋白基因工程表达载体的构建  

Constrction of a Bivalent Expression Vector of Outer Membrane Protein of Two Pathogens Pathogenic to Eels

作　　者：郭松林[1,2] 王玉[1,2] 冯建军[1,2] 林鹏[1,2] 

机构地区：[1]集美大学水产学院,福建厦门361021 [2]鳗鲡现代产业技术教育部工程研究中心,福建厦门361021

出　　处：《集美大学学报（自然科学版）》2014年第5期330-338,共9页Journal of Jimei University：Natural Science

基　　金：国家自然科学基金资助项目(31001136)

摘　　要：根据鳗鲡病原性嗜水气单胞菌Ⅱ型孔蛋白(porinⅡ)和迟钝爱德华氏菌外膜蛋白S(ompS2)的基因全长序列,分别选取这两个全长序列中表达其蛋白质膜外部分且理论免疫原性较好的两个基因片段,通过融合PCR技术连接这两个外膜蛋白基因片段.根据表达载体(pGEX-2T-His)的限制性酶切位点在连接序列片段的两端引入限制性酶切位点BamHⅠ和EcoRⅠ,成功构建了双外膜蛋白基因片段重组表达载体(pGEX-2T-His-porinⅡ-ompS2).表达载体理论表达产物的蛋白质结构预测表明表达产物无信号肽和跨膜区,不存在三级结构;亲水性和免疫原性预测分析表明该表达蛋白理论上为可溶性蛋白并具有丰富的抗原决定簇,本研究为该表达载体的蛋白表达、纯化以及表达产物的免疫原性研究奠定了基础.A fusion DNA that composed of fragments of two genes that,encoding outer region of the outer termembrane proteins which have good immunogenicity of Aeromonas hydrophila（ porin Ⅱ） and Edwardsiella tarda（ ompS2）,were obtained by the＂two-steps＂fusion PCR. Cut sites of BamHⅠ and EcoRⅠ were added at two 5＇end of two primers amplifying the fusion DNA respectively,The recombinant expression vector（ pGEX-2T-His-porin Ⅱ-ompS2） expressing fused protein was constructed. Protein structure prediction showed that the fused protein（ bivalent outer membrane protein） had no signal peptide,transmembrane region or three stage structure. Hydrophilic and immunogenicity analysis showed that the expressed fused protein is a soluble protein that is made up of many antigenic determinants. This study laid a foundation for the expression,purification and immunogenicity study of the protein expressed by the recombinant expression vector.

关 键 词：嗜水气单胞菌 迟钝爱德华氏菌 鳗鲡 二联外膜蛋白 表达载体 

分 类 号：S942.5[农业科学—水产养殖]