發(fā)布時間：2018-01-16 10:02

  本文關鍵詞：幽門螺桿菌對阿莫西林耐藥機制的初步探索　出處：《山西醫(yī)科大學》2007年碩士論文　論文類型：學位論文

  更多相關文章： 幽門螺桿菌 抗性 蛋白質(zhì)組 雙向電泳 阿莫西林

【摘要】： 目的通過體外誘導產(chǎn)生幽門螺桿菌對阿莫西林耐藥菌株,對耐藥菌株和敏感菌株表達差異蛋白點的功能進行鑒定。 方法以J99為出發(fā)菌株,采用連續(xù)傳代法,將J99在含有AMX的抗性平板上連續(xù)傳代,獲得耐藥菌株,采用三氯醋酸/丙酮沉淀法制取敏感菌株和耐藥菌株的全菌蛋白,對耐藥菌株和敏感菌株做雙向凝膠電泳,通過Image Master 2D Elite軟件對其蛋白質(zhì)圖譜進行比較,從而找到表達差異的蛋白,對表達差異的蛋白點進行膠內(nèi)酶解,利用質(zhì)譜技術和生物信息學分析差異蛋白的性質(zhì)和功能。 結果通過質(zhì)譜和數(shù)據(jù)庫搜索對這些表達差異的蛋白點進行鑒定顯示8個表達差異的蛋白斑點,其中1個蛋白斑點S-adenosylmethionine synthetase只在AMX原始菌株中表達,1個蛋白斑點co-chaperonin GroES只在耐藥菌株中表達。2個蛋白斑點30S ribosomal proteinS4和flagellin在耐藥菌株中表達上調(diào),4個蛋白斑點2-oxoglutarate ferredoxinoxidoreductase、3-OXOACID COA-TRANSFERASE,SUBUNIT A、NADH dehydrogenasesubunitⅠ和hypothetical protein在敏感菌株中表達量高于耐藥菌株。 結論通過體外誘導產(chǎn)生一株耐阿莫西林菌株,當有效的藥物作用于細菌時,細菌的代謝功能發(fā)生變化,導致細菌表達的蛋白發(fā)生改變,對這些差異表達蛋白的研究有助于了解細菌耐藥機制的形成。
[Abstract]:Objective to induce Helicobacter pylori to amoxicillin resistant strains in vitro and to identify the function of differentially expressed protein spots between resistant strains and sensitive strains. Methods using J99 as the starting strain, J99 was subcultured on the resistant plate containing AMX by continuous passage method, and the drug-resistant strain was obtained. The whole bacterial proteins of sensitive and resistant strains were prepared by dichloroacetic acid / acetone precipitation method, and two dimensional gel electrophoresis was carried out on resistant and sensitive strains. The protein map was compared by Image Master 2D Elite software, and the differentially expressed proteins were found, and the differentially expressed protein spots were hydrolyzed in the gel. The properties and functions of differential proteins were analyzed by mass spectrometry and bioinformatics. Results eight differentially expressed protein spots were identified by mass spectrometry and database search. One protein spot, S-adenosylmethionine synthetase, was expressed only in the original AMX strain. 1 protein spot co-chaperonin GroES was expressed only in resistant strains. 2 protein spots 30s ribosomal. The expression of proteinS4 and flagellin was up-regulated in drug-resistant strains. Four protein spots 2-oxoglutarate ferredoxinoxidoreductase. 3-OXOACID COA-TRANSFERASE / SUBUNIT. The expression of NADH dehydrogenasesubunit 鈪，

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