解甘露醇罗尔斯顿菌金属蛋白酶基因特征分析  

Analysis on characteristics of metalloprotease genes in Ralstonia mannitolilytica strain

作　　者：郁文燕 徐爱芳 潘克女 葛玉梅[2] 

机构地区：[1]杭州市西溪医院检验科,310023 [2]浙江省人民医院检验中心,杭州310014

出　　处：《国际流行病学传染病学杂志》2017年第3期161-166,共6页International Journal of Epidemiology and Infectious Disease

基　　金：浙江省医药卫生科技计划（2017KY004）

摘　　要：目的检测解甘露醇罗尔斯顿菌临床分离株的金属蛋白酶基因，并分析其编码蛋白结构特征。方法采用Axygen细菌基因组提取试剂盒提取解甘露醇罗尔斯顿菌临床分离株DNA，建立基于金属蛋白酶基因的PCR检测方法，使用BLAST比对基因序列，使用CDD、MEROPS、SWISS．MODEL、CDTree、TMHMM和SignalP4．1对编码蛋白进行在线分析。结果在该解甘露醇罗尔斯顿菌临床分离株中分别检测出M48和S2P-M50家族金属蛋白酶基因RS—MP48和RS-MP50，与已报道的解甘露醇罗尔斯顿菌SN82F48株所携带的基因序列（WP-045785189．1和WP-045786756．1）相似度为100％。生物信息学研究表明，RS．MP48含有1个M48蛋白酶超家族保守功能域和4个跨膜结构，与芽孢杆菌M48肽酶家族、芽孢杆菌锌依赖蛋白酶、铜绿假单胞菌锌依赖蛋白酶同源性最高，相似度均为99％；与链霉菌热休克蛋白和铜绿微囊藻蛋白酶相似度分别为33％和32％；RS-MP50含有2个S2P—M50超家族保守功能域和4个明显的跨膜结构及少量的膜结合区；与解甘露醇罗尔斯顿菌RIP金属蛋白酶RseP同源性最高，相似度为99％，与罗尔斯顿菌属RIP金属蛋白酶RseP和蛋白酶调节因子相似度均为92％，与脑膜炎奈瑟菌和希瓦菌RseA锌金属蛋白降解酶RseP相似度分别为39％和38％。结论解甘露醇罗尔斯顿菌是引起人类感染的新病原菌，本研究发现该临床株中存在金属蛋白酶基因，其编码蛋白可能是该类细菌潜在的毒力因子。Objective To detect the metalloprotease genes in the clinical isolates of R. mannitolilytica, and analyze the structural characteristics of the encoding proteins. Methods DNA from clinical isolates of R. mannitolilytica was extracted by Axygen bacterial genome extraction kit. Metalloprotease genes-based PCR detection method was established. BLAST was used to compare the gene sequence. CDD, MEROPS, SWISS-MODEL, CDTree, TMHMM and SignalP 4.1 were used to analyze encoding proteins online. Results The R. mannitolilytica clinical strain contained RS-MP48 gene and RS-MPS0 gene of M48 superfamily and S2P-MS0 superfamily, respectively. The genes were consistent with the reported genes （WP_045785189.1 and WP_045786756.1） in R. mannitolilytica strain SN82F48 （identity=100%）. Bioinformatics research showed that RS-MP48 contained one conserved domain of M48 metalloprotease superfamily and four transmembrane structures, and had highest homology with M48 metalloprotease, Zinc dependent protease of Methylobacillus flagellatus KT and Zinc dependent protease of Pseudomonas aeruginosa with 99% identity; while it only had 33% homology with heat shock protein in Streptomyces coelicolor and 32% homology with protease in Microcystis aeruginosca RS-MPS0 contained two conserved domains of S2P-MS0 superfamily and four transmembrane structures with several membrane binding regions, and had highest homology with RseP in R. mannitolilytica with 99% identity; while it had 92% homology with RseP in Ralstonia sp. A12 and protease regulator in Ralstoniasp. NFACC01, as well as only 39% homology with NMB0183 protein in Neisseria meningitidis MC58 and 38% homology with Zinc dependent degrading enzyme in Shewanella oneidensis MR-1. Conclusions R. mannitolilytica is an emerging bacterial pathogen that leads to human infectious diseases. In this study, there is a metalloprotease gene in the clinical isolate of R. mannitolilytica and its encoded protein may be a potential virulence factor.

关 键 词：金属蛋白酶类 解甘露醇罗尔斯顿菌 基因特征 生物信息学 

分 类 号：R378[医药卫生—病原生物学]