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核酸适配体对生物膜形成的有效抑制作用

Efficient suppression of biofilm formation by a nucleic acid aptamer.

作者信息

Ning Yi, Cheng Lijuan, Ling Min, Feng Xinru, Chen Lingli, Wu Minxi, Deng Le

机构信息

Department of Microbiology, College of Life Science, Hunan Normal University, Changsha, Hunan 410081, People's Republic of China The Medicine School of Hunan University of Chinese Medicine, Changsha, Hunan 410208, People's Republic of China.

Department of Microbiology, College of Life Science, Hunan Normal University, Changsha, Hunan 410081, People's Republic of China.

出版信息

Pathog Dis. 2015 Aug;73(6):ftv034. doi: 10.1093/femspd/ftv034. Epub 2015 May 29.

Abstract

Biofilms are microbial communities that are attached to a solid surface using extracellular polymeric substances. Motility and initial attachment mediated by flagella are required for biofilm formation. Therefore, blocking the motility of flagella is a potential strategy to inhibit biofilm formation. In this study, single-stranded DNA aptamers specific to the Salmonella choleraesuis were selected after 14 cycles of the systematic evolution of ligands by exponential enrichment. Among the selected aptamers, the aptamer 3 showed the highest affinity for S. choleraesuis with a dissociation constant (Kd) of 41 ± 2 nM. Aptamer 3, conjugated with magnetic beads, was then used to capture its binding target on the bacteria. After mass spectrometry and specific binding analysis, the flagellin was identified as the target captured by aptamer 3. Furthermore, inhibition experiments, inverted microscopy and atomic force microscopy demonstrated that aptamer 3 was able to control the biofilm formation and promote the inhibitory effect of an antibiotic on bacterial biofilms. Single-stranded DNA aptamers therefore have great potential as inhibitors of biofilm formation.

摘要

生物膜是利用细胞外聚合物附着于固体表面的微生物群落。生物膜形成需要由鞭毛介导的运动性和初始附着。因此,阻断鞭毛的运动性是抑制生物膜形成的一种潜在策略。在本研究中,通过指数富集配体系统进化经过14轮筛选出了特异性针对猪霍乱沙门氏菌的单链DNA适配体。在所选的适配体中,适配体3对猪霍乱沙门氏菌表现出最高亲和力,解离常数(Kd)为41±2 nM。然后将与磁珠偶联的适配体3用于捕获其在细菌上的结合靶点。经过质谱分析和特异性结合分析,确定鞭毛蛋白是被适配体3捕获的靶点。此外,抑制实验、倒置显微镜和原子力显微镜表明,适配体3能够控制生物膜的形成,并增强抗生素对细菌生物膜的抑制作用。因此,单链DNA适配体作为生物膜形成抑制剂具有巨大潜力。

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