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

  更多相關(guān)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(dǎo)產(chǎn)生幽門螺桿菌對(duì)阿莫西林耐藥菌株,對(duì)耐藥菌株和敏感菌株表達(dá)差異蛋白點(diǎn)的功能進(jìn)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(duì)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(duì)其蛋白質(zhì)圖譜進(jìn)行比較,從而找到表達(dá)差異的蛋白,對(duì)表達(dá)差異的蛋白點(diǎn)進(jìn)行膠內(nèi)酶解,利用質(zhì)譜技術(shù)和生物信息學(xué)分析差異蛋白的性質(zhì)和功能。 結(jié)果通過質(zhì)譜和數(shù)據(jù)庫搜索對(duì)這些表達(dá)差異的蛋白點(diǎn)進(jìn)行鑒定顯示8個(gè)表達(dá)差異的蛋白斑點(diǎn),其中1個(gè)蛋白斑點(diǎn)S-adenosylmethionine synthetase只在AMX原始菌株中表達(dá),1個(gè)蛋白斑點(diǎn)co-chaperonin GroES只在耐藥菌株中表達(dá)。2個(gè)蛋白斑點(diǎn)30S ribosomal proteinS4和flagellin在耐藥菌株中表達(dá)上調(diào),4個(gè)蛋白斑點(diǎn)2-oxoglutarate ferredoxinoxidoreductase、3-OXOACID COA-TRANSFERASE,SUBUNIT A、NADH dehydrogenasesubunitⅠ和hypothetical protein在敏感菌株中表達(dá)量高于耐藥菌株。 結(jié)論通過體外誘導(dǎo)產(chǎn)生一株耐阿莫西林菌株,當(dāng)有效的藥物作用于細(xì)菌時(shí),細(xì)菌的代謝功能發(fā)生變化,導(dǎo)致細(xì)菌表達(dá)的蛋白發(fā)生改變,對(duì)這些差異表達(dá)蛋白的研究有助于了解細(xì)菌耐藥機(jī)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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