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Title: | 霍亂沙門氏桿菌分離株之比較蛋白質體分析 Comparative Proteomics Analysis of Salmonella enterica serovar Choleraesuis Isolates |
Authors: | Chih-Kuan Chen 陳志冠 |
Advisor: | 何國傑(Kuo-Chieh Ho) |
Keyword: | 霍亂沙門氏桿菌,抗藥性,寄主性,二維電泳,即時偵測定量聚合脢連鎖反應,獨立演化, Salmonella Choleraesuis,Drug resistance,Host specificity,Real Time QPCR,Isolate evolution, |
Publication Year : | 2007 |
Degree: | 碩士 |
Abstract: | 霍亂沙門氏桿菌(Salmonella Choleraesuis)是豬隻主要的致病菌之一,偶爾可傳染給人類,並造成系統性的感染。過去多以抗生素來施予治療,但在近年來的研究卻發現,霍亂沙門氏桿菌在台灣等地區出現越來越高的抗藥性比率,尤其對flouroquinolone(FQ)類藥物的抗性正漸漸產生,且遭受感染的人數也逐漸增加中。目前對於霍亂沙門氏桿菌的研究主要以流行病學的基因分子分型為主,較少有基因或蛋白質表現層次的研究。本實驗自疾病管制局從台灣各地所分離的199隻菌種中,挑選了四種不同的分離株,分別是抗FQ的人體分離株,不抗FQ的人體分離株,以及抗FQ的豬隻分離株,不抗FQ的豬隻分離株。試圖以蛋白質體學的方式,對這些分離株進行交叉比對,以找出高低抗藥性與不同寄主間的差異蛋白。具有差異表現的蛋白經過鑑定後,再以QPCR確認這些蛋白在基因表現層次的差異,總共篩選出14個蛋白質,分別是與抗FQ相關的SC064, sat, dps, groS, groL, pduB, malE;與寄主差異相關的sat, dps, groL, pduB, menB, STY3221;在豬與人分離株對FQ抗性有不同反應的基因uspA, pgk, dnaK, rpsA, tuf A,B。這些基因顯示:霍亂沙門氏桿菌對於逆境可能有廣泛性的適應機制,利用dps與熱休克蛋白的調控來適應FQ以及新寄主(人體)。菌體在人體內遭受的逆境可能較在豬隻體內更為強烈,因此有調降生長發育相關的基因來維持生存的情形。菌體在不同寄主體內,可能對抗藥性出現獨立演化的情形,因此人體分離株的抗藥性有普遍較高的現象。 Salmonella Choleraesuis is one of the most important pathogens for pigs. It occasionally infects human and causes serious systemic infection. Although antibiotics are the effective drug to prevent and cure the disease, evidences suggest that more and more isolates of S. Choleraesuis in Taiwan have developed drug resistance, especially to flouroquinolone (FQ), and the cases of human infection are increasing. In the last few years, studies have been devoted to genotyping of S. Choleraesuis for epidemiology, but only few have been made at the expression patterns of genes or proteins of S. Choleraesuis. In this study, the proteomics analysis was performed to determine the differential patterns of proteins. Four different isolates were chosen from 199 isolates isolated from Taiwan by Centers for Disease Control: one FQ resistant isolate and one FQ susceptible isolate from human, and one FQ resistant isolate and one FQ susceptible isolate from pig. After the differential displayed proteins were identified by 2D-electrophoresis, the expression level of individual protein was confirmed by Real Time QPCR. Finally, fourteen candidate proteins were identified. Of which, seven proteins could be associated with FQ resistance: SC064, sat, dps, groS, groL, pduB, malE, six proteins could be related to host specificity: sat, dps, groL, pduB, menB, STY3221, and five proteins could be related to FQ resistance in different hosts:sat, dps, groL, pduB, menB, STY3221. The results suggested that S. Choleraesuis could possess universal mechanisms in adaptation to FQ and host by regulating the expression of dps and HSPs. S. Choleraesuis might suffer more stress within human than in pigs, as the genes related to growth and development were down-regulated. S. Choleraesuis might have different drug resistant within different hosts via isolated evolution, therefore human isolates appear to have higher drug resistance. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/30955 |
Fulltext Rights: | 有償授權 |
Appears in Collections: | 植物科學研究所 |
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