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军团菌激活的 RNA 切割 DNA 酶的选择与鉴定。

Selection and Characterization of an RNA-Cleaving DNAzyme Activated by Legionella pneumophila.

机构信息

Michael G. DeGroote Institute for Infectious Disease Research, Department of Biochemistry and Biomedical Sciences, McMaster University, 1280 Main Street West, Hamilton, ON, L8S 4K1, Canada.

Department of Biochemistry, University of Toronto, Canada.

出版信息

Angew Chem Int Ed Engl. 2021 Feb 23;60(9):4782-4788. doi: 10.1002/anie.202012444. Epub 2021 Jan 15.

Abstract

Legionella pneumophila is a deadly bacterial pathogen that has caused numerous Legionnaires' disease outbreaks, where cooling towers were the most common source of exposure. Bacterial culturing is used for L. pneumophila detection, but this method takes approximately 10 days to complete. In this work, an RNA-cleaving fluorogenic DNAzyme, named LP1, was isolated. Extensive characterization revealed that LP1 is reactive with multiple infectious isolates of L. pneumophila but inactive with 25 other common bacterial species. LP1 is likely activated by a protein target, capable of generating a detectable signal in the presence of as few as 10 colony-forming units of L. pneumophila, and able to maintain its activity in cooling tower water from diverse sources. Given that similar DNAzymes have been incorporated into many sensitive assays for bacterial detection, LP1 holds the potential for the development of biosensors for monitoring the contamination of L. pneumophila in exposure sources.

摘要

嗜肺军团菌是一种致命的细菌病原体,已导致多次军团病暴发,其中冷却塔是最常见的暴露源。细菌培养用于检测嗜肺军团菌,但这种方法需要大约 10 天才能完成。在这项工作中,分离出了一种名为 LP1 的 RNA 切割荧光 DNA 酶。广泛的表征表明,LP1 与多种传染性嗜肺军团菌分离株反应,但对 25 种其他常见细菌无活性。LP1 可能被一种蛋白质靶标激活,在存在多达 10 个嗜肺军团菌集落形成单位的情况下,能够产生可检测的信号,并能够在来自不同来源的冷却塔水中保持其活性。鉴于类似的 DNA 酶已被纳入许多用于细菌检测的敏感检测中,LP1 有可能开发出用于监测暴露源中嗜肺军团菌污染的生物传感器。

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