迟钝爱德华氏菌Ⅲ型分泌系统效应蛋白功能域预测  

Prediction of Functional Domain of Effector Protein of TypeⅢ Secretion System in Pathogen Edwardsiella tarda

作　　者：王斌[1] 李军伟[1] 赵凤梅[1] 王惠[1] 胡亮[1] 姜志强[1] 

机构地区：[1]大连海洋大学,农业部北方海水增养殖重点实验室,辽宁省高校海洋生物资源可持续利用重点实验室,辽宁大连116023

出　　处：《水产科学》2013年第7期385-390,共6页Fisheries Science

基　　金：辽宁省教育厅科学技术研究重点实验室项目(2009S026);辽宁省重大科技计划项目(2011203005)

摘　　要：采用PCR法检测了大菱鲆出血性败血症病原菌迟钝爱德华氏菌L-49231菌株Ⅲ型分泌系统中的eseB、eseC、eseD基因,经PCR产物克隆测序;利用生物信息学方法对eseB、eseC、eseD表达蛋白的分子量、氨基酸组成、疏水性、跨膜结构、信号肽等进行初步预测和分析。结果显示,迟钝爱德华氏菌L-49231菌株eseB、eseC、eseD基因分别为:594、934、445bp,与GenBank中登录的迟钝爱德华氏菌PPD FL6-60菌株的eseB、eseC、eseD基因碱基序列的同源性分别为100%、100%、99%。eseB蛋白分子量为21 732.26u,天冬氨酸含量最高,占10.61%;eseC蛋白分子量为53 606.08u,丙氨酸含量最高,占14.43%;eseD蛋白的分子量为21 112.43u,谷氨酰胺含量最高,占11.04%。3种蛋白质中甘氨酸、丙氨酸、亮氨酸含量都很高,而半胱氨酸、色氨酸含量都很低或缺少。3种蛋白均为亲水性蛋白质。eseB无明显的跨膜结构;eseC在192～210aa、213～233aa、263～284aa处形成3个跨膜区域;eseD在90～106aa处形成一个跨膜区域。3种蛋白均不含信号肽。The PCR was performed to detect the TTSS genes eseB, eseC, and eseD in pathogen Edward- siella tarda L-49231 strain isolated from turbot Scophthalmus maximus infected with septicaemia hemor- rhage. The PCR products were cloned into vector PMD19-T and the positive clones were sequenced. The results showed that the fragments were composed of 594 bp in eseB, 934 bp in eseC, and 445 bp in eseD, showing 100%, 100%, and 99% homology separately compared to E. tarda PPD FL6-60 in GenBank. Thereby, the E. tarda L-49231 strain contained the TTSS effector genes eseB, eseC, and ese D. The molecular weight, amino acid composition, hydrophobicity, transmembrane structure and the signal peptide of eseB, eseC and eseD were predicted by bioinformatics. The eseB was found to be 21 732.26 u, with the maximal level of aspartic acid, representing 10. 61% of the total amino acids. The eseC had molecular weight of 53 606.08 u, with maximal level of alanine, accounting for 14.43% of the total amino acids. The eseD showed the molecular weight of 21 112.43 u, glutamine being the maximum, representing 11.04%of the total amino acids. In addition, all of the three proteins contained much glycin, alanine and leucine but a little eysteine and tryptophan. The eseB, eseC and eseD were all found to be hydrophilic proteins, without transmembrane structure in the eseB. There was three transmembrane structures at 192-210 aa, 213-233 aa and 63-284 aa domain in eseC. The eseD contained a transmembrane structure at 90-106 aa domain. There was no signal peptide in all of the three proteins.

关 键 词：迟钝爱德华氏菌 Ⅲ型分泌系统 克隆 生物信息 

分 类 号：S941[农业科学—水产养殖]