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一种来自鸟分枝杆菌的新型抗氧化基因对大肠杆菌亚硝化和氧化应激的保护作用。

The protective effect of a novel antioxidant gene from Mycobacterium avium against nitrosative and oxidative stress in E. coli.

作者信息

He Shiyi, Wang Aiyan, Ning Xueping, Yang Dongjun, Ling Min

机构信息

Key Laboratory of Biological Molecular Medicine Research of Guangxi University, Department of Biochemistry and Molecular Biology, Guangxi Medical University, Nanning, 530021, People's Republic of China.

Department of Clinical Laboratory, Second Affiliated Hospital of Guangxi Medical University, Nanning, 530007, People's Republic of China.

出版信息

World J Microbiol Biotechnol. 2017 Jun;33(6):127. doi: 10.1007/s11274-017-2294-6. Epub 2017 May 25.

Abstract

The production of reactive oxygen intermediates (ROI) and reactive nitrogen intermediates (RNI) is an important host defense mechanism in response to infection by Mycobacterium tuberculosis. A variety of genes have been implicated in resistance to ROI and RNI, including noxR1. However, studies in Mycobacterium avium, an important pathogen among nontuberculous mycobacteria, are limited. We aim to investigate the role of a novel gene cloned from M. avium with high similarity to noxR1, noA, in resistance against RNI and ROI in M. tuberculosis. After subcloning noA into vector for expression in E. coli, we performed survival rate analysis in the bacteria transformed with noA (pET-noA) and without noA (pET-his) after exposure to nitrosative stresses by S-nitrosoglutathione (GSNO) and sodium nitrite, and oxidative stresses by HO. Compared with pET-his, the survival rate of pET-noA was 1 log-fold higher after exposure to GSNO and sodium nitrite. We observed 1 log-fold, 2 log-fold and 3 log-fold higher survival rate in pET-noA than pET-his after exposure to HO for 3, 6 and 9 h, respectively. With the combined treatment of HO and GSNO, we found more than 2 log-fold increase in survival rate in pET-noA comparing with pET-his, suggesting a possible synergistic effect. In summary, noA gene cloned from M. avium has been shown to protect E. coli from both RNI and ROI.

摘要

活性氧中间体(ROI)和活性氮中间体(RNI)的产生是宿主针对结核分枝杆菌感染的一种重要防御机制。多种基因与对ROI和RNI的抗性有关,包括noxR1。然而,对非结核分枝杆菌中的重要病原体鸟分枝杆菌的研究有限。我们旨在研究从鸟分枝杆菌克隆的与noxR1高度相似的新基因noA在结核分枝杆菌对RNI和ROI抗性中的作用。将noA亚克隆到载体中以便在大肠杆菌中表达后,我们对用noA(pET-noA)和未用noA(pET-his)转化的细菌进行了存活率分析,这些细菌在暴露于由S-亚硝基谷胱甘肽(GSNO)和亚硝酸钠引起的亚硝化应激以及由HO引起的氧化应激后。与pET-his相比,暴露于GSNO和亚硝酸钠后,pET-noA的存活率高1个对数级。暴露于HO 3、6和9小时后,我们观察到pET-noA的存活率分别比pET-his高1个对数级、2个对数级和3个对数级。在用HO和GSNO联合处理时,我们发现与pET-his相比,pET-noA的存活率增加超过2个对数级,表明可能存在协同效应。总之,从鸟分枝杆菌克隆的noA基因已被证明可保护大肠杆菌免受RNI和ROI的影响。

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