幽门螺杆菌对阿莫西林耐药机制的初步探索

发布时间：2018-01-16 10:02

  本文关键词：幽门螺杆菌对阿莫西林耐药机制的初步探索　出处：《山西医科大学》2007年硕士论文　论文类型：学位论文

  更多相关文章： 幽门螺杆菌 抗性 蛋白质组 双向电泳 阿莫西林

【摘要】： 目的通过体外诱导产生幽门螺杆菌对阿莫西林耐药菌株,对耐药菌株和敏感菌株表达差异蛋白点的功能进行鉴定。 方法以J99为出发菌株,采用连续传代法,将J99在含有AMX的抗性平板上连续传代,获得耐药菌株,采用三氯醋酸/丙酮沉淀法制取敏感菌株和耐药菌株的全菌蛋白,对耐药菌株和敏感菌株做双向凝胶电泳,通过Image Master 2D Elite软件对其蛋白质图谱进行比较,从而找到表达差异的蛋白,对表达差异的蛋白点进行胶内酶解,利用质谱技术和生物信息学分析差异蛋白的性质和功能。 结果通过质谱和数据库搜索对这些表达差异的蛋白点进行鉴定显示8个表达差异的蛋白斑点,其中1个蛋白斑点S-adenosylmethionine synthetase只在AMX原始菌株中表达,1个蛋白斑点co-chaperonin GroES只在耐药菌株中表达。2个蛋白斑点30S ribosomal proteinS4和flagellin在耐药菌株中表达上调,4个蛋白斑点2-oxoglutarate ferredoxinoxidoreductase、3-OXOACID COA-TRANSFERASE,SUBUNIT A、NADH dehydrogenasesubunitⅠ和hypothetical protein在敏感菌株中表达量高于耐药菌株。 结论通过体外诱导产生一株耐阿莫西林菌株,当有效的药物作用于细菌时,细菌的代谢功能发生变化,导致细菌表达的蛋白发生改变,对这些差异表达蛋白的研究有助于了解细菌耐药机制的形成。
[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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