本文選題：β-內(nèi)酰胺抗生素 + 細菌耐藥性��； 參考：《華東理工大學》2017年碩士論文

【摘要】：β-內(nèi)酰胺類抗生素是目前治療感染類疾病最為重要的藥物之一,為人類的健康生活提供了重要的醫(yī)療保障。但是,隨著這類抗生素的大量應用,臨床上出現(xiàn)了越來越多的耐藥菌,導致抗生素的藥效大大降低,甚至出現(xiàn)了幾乎能夠抵抗所有抗生素的"超級細菌"。病菌產(chǎn)生耐藥性的一個主要原因是細菌產(chǎn)生了一種能夠水解破壞β-內(nèi)釀胺類抗生素的酶,即β-內(nèi)酰胺酶�，F(xiàn)有研究表明,β-內(nèi)酰胺類抗生素與相應的β-內(nèi)酰胺酶抑制劑聯(lián)合使用能夠很大程度地恢復抗生素的抑菌效力。然而,對于金屬主導的一類β-內(nèi)酰胺酶目前還沒有能夠在臨床中應用的β-內(nèi)酰胺酶抑制劑,因此對于β-內(nèi)酰胺酶抑制劑的研究仍然是當前非常重要的一個任務。目前β-內(nèi)酰胺酶抑制劑的篩選主要是利用底物水解前后紫外吸收的差異,該方法所需底物量較大,紫外吸收波長短,易受到外界的干擾。在本論文中,我們通過對β-內(nèi)酰胺酶基因的載體進行篩選和蛋白表達條件進行優(yōu)化,最終完成了 8個不同類型的β-內(nèi)酰胺酶的表達純化,并應用部分β-內(nèi)酰胺酶對熒光探針的水解進行了系統(tǒng)研究,通過相關條件優(yōu)化完成了 β-內(nèi)酰胺酶抑制劑快速篩選平臺的構建。本論文建立基于β-內(nèi)酰胺酶熒光探針的β-內(nèi)酰胺酶抑制劑快速篩選平臺,該平臺利用熒光探針水解前后熒光強度的變化檢測酶的水解活性,具有檢測靈敏度高,酶的使用量少等優(yōu)點,尤其適用于對潛在活性化合物的高通量篩選。
[Abstract]:尾-lactam antibiotics are currently one of the most important drugs for the treatment of infectious diseases, which provide an important medical guarantee for the healthy life of human beings. However, with the extensive application of these antibiotics, more and more drug-resistant bacteria appear in clinic, which leads to the decrease of antibiotic efficacy and even the emergence of "super bacteria" which can resist almost all antibiotics. One of the main reasons for bacterial resistance is that bacteria produce an enzyme that can hydrolyze and destroy 尾-lactamases, or 尾-lactamases. Current studies have shown that the combination of 尾-lactam antibiotics and corresponding 尾-lactamases inhibitors can restore the antimicrobial efficacy of antibiotics to a large extent. However, there is no 尾 -lactamase inhibitor that can be used in clinic, so the research of 尾 -lactamase inhibitor is still a very important task. At present, the screening of 尾 -lactamase inhibitors is mainly based on the difference of UV absorption before and after substrate hydrolysis. In this thesis, we selected the vector of 尾 -lactamase gene and optimized the expression conditions of 尾 -lactamases, and finally completed the purification of eight different types of 尾 -lactamases. Some 尾 -lactamases were used to study the hydrolysis of fluorescent probes, and the rapid screening platform of 尾 -lactamase inhibitors was constructed by optimizing the relevant conditions. In this paper, a rapid screening platform for 尾 -lactamase inhibitors based on fluorescence probe of 尾 -lactamase was established. The platform used fluorescence probe to detect the activity of 尾 -lactamase by using fluorescence probe before and after hydrolysis. It is especially suitable for high throughput screening of potential active compounds.
【學位授予單位】：華東理工大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R91

【參考文獻】

相關期刊論文 前1條

1 陶虹;盧體康;唐金明;廖立珊;阮周曦;劉建利;曾少靈;陳兵;胡運發(fā);孫潔;曹琛福;張彩虹;呂建強;楊俊興;花群義;秦智鋒;;產(chǎn)CTX-M和TEM型ESBLs耐藥大腸埃希菌雙重PCR檢測方法的建立[J];動物醫(yī)學進展;2013年08期

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