Genetic Manipulation of Corynebacterium diphtheriae and Other Corynebacterium Species

Current Protocols in Microbiology Pub Date : 2020-08-31 DOI:10.1002/cpmc.111

Chungyu Chang, Minh Tan Nguyen, Hung Ton-That

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引用次数: 2

Abstract

This article describes several established approaches for genetic manipulation of Corynebacterium diphtheriae, the causative agent of diphtheria that is known to have provided key evidence for Koch's postulates on the germ theory. First, it includes a detailed gene deletion method that generates nonpolar, in-frame, markerless deletion mutants, utilizing the levansucrase SacB as a counter-selectable marker. Second, it provides a thorough protocol for rescuing deletion mutants using Escherichia coli–Corynebacterium shuttle vectors. Finally, a Tn5 transposon mutagenesis procedure is described. In principle, these protocols can be used for other Corynebacterium species, including Corynebacterium glutamicum and Corynebacterium matruchotii. © 2020 Wiley Periodicals LLC

Basic Protocol 1: Gene deletion in Corynebacterium diphtheriae

Basic Protocol 2: Complementation of a mutant strain

Basic Protocol 3: Tn5 transposon mutagenesis of Corynebacterium diphtheriae

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白喉棒状杆菌和其他棒状杆菌的遗传操作

本文描述了白喉棒状杆菌遗传操作的几种既定方法，白喉棒状杆菌是白喉的病原体，已知它为科赫在细菌理论上的假设提供了关键证据。首先，它包括一种详细的基因缺失方法，该方法利用左旋蔗糖酶SacB作为反选择标记，产生非极性、框架内、无标记的缺失突变体。其次，它为利用大肠杆菌-棒状杆菌穿梭载体挽救缺失突变体提供了一个完整的方案。最后，描述了Tn5转座子的突变过程。原则上，这些方案可用于其他棒状杆菌种类，包括谷氨酸棒状杆菌和matruchotii棒状杆菌。©2020 Wiley期刊llc基本方案1:白喉棒状杆菌基因缺失基本方案2:突变菌株的互补基本方案3:白喉棒状杆菌Tn5转座突变

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Current Protocols in Microbiology Immunology and Microbiology-Parasitology

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期刊介绍： Current Protocols in Microbiology provides detailed, step-by-step instructions for analyzing bacteria, animal and plant viruses, fungi, protozoans and other microbes. It offers updated coverage of emerging technologies and concepts, such as biofilms, quorum sensing and quantitative PCR, as well as proteomic and genomic methods. It is the first comprehensive source of high-quality microbiology protocols that reflects and incorporates the new mandates and capabilities of this robust and rapidly evolving discipline.

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