Improved methods for genetic manipulation of the alkaliphile Halalkalibacterium halodurans

IF 4 2区生物学 Q2 MICROBIOLOGY

Frontiers in Microbiology Pub Date : 2024-09-18 DOI:10.3389/fmicb.2024.1465811

Freya D. R. Wencker, Seth E. Lyon, Ronald R. Breaker

引用次数: 0

Abstract

An improved approach was developed for the genetic manipulation of the Gram-positive extremophile Halalkalibacterium halodurans (formerly called Bacillus halodurans). We describe an allelic replacement method originally developed for Staphylococcus aureus that allows the deletion, mutation, or insertion of genes without leaving markers or other genetic scars. In addition, a protocol for rapid in vitro plasmid methylation and transformation is presented. The combined methods allow the routine genetic manipulation of H. halodurans from initial transformation to the desired strain in 8 days. These methods improve H. halodurans as a model organism for the study of extremophiles.

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卤化嗜碱杆菌遗传操作方法的改进

我们开发了一种改进的方法来对革兰氏阳性极端嗜酸性杆菌 Halalkalibacterium halodurans（原名 Halodurans 杆菌）进行遗传操作。我们描述了一种等位基因置换方法，这种方法最初是为金黄色葡萄球菌开发的，可以在不留下标记或其他遗传疤痕的情况下删除、突变或插入基因。此外，还介绍了一种快速体外质粒甲基化和转化的方案。通过这些综合方法，可在 8 天内对卤虫进行从初始转化到所需菌株的常规遗传操作。这些方法改进了哈洛杜兰菌，使其成为研究嗜极生物的模式生物。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Frontiers in Microbiology MICROBIOLOGY-

CiteScore

7.70

自引率

9.60%

发文量

4837

审稿时长

14 weeks

期刊介绍： Frontiers in Microbiology is a leading journal in its field, publishing rigorously peer-reviewed research across the entire spectrum of microbiology. Field Chief Editor Martin G. Klotz at Washington State University is supported by an outstanding Editorial Board of international researchers. This multidisciplinary open-access journal is at the forefront of disseminating and communicating scientific knowledge and impactful discoveries to researchers, academics, clinicians and the public worldwide.