Freya D. R. Wencker, Seth E. Lyon, Ronald R. Breaker
{"title":"Improved methods for genetic manipulation of the alkaliphile Halalkalibacterium halodurans","authors":"Freya D. R. Wencker, Seth E. Lyon, Ronald R. Breaker","doi":"10.3389/fmicb.2024.1465811","DOIUrl":null,"url":null,"abstract":"An improved approach was developed for the genetic manipulation of the Gram-positive extremophile <jats:italic>Halalkalibacterium halodurans</jats:italic> (formerly called <jats:italic>Bacillus halodurans</jats:italic>). We describe an allelic replacement method originally developed for <jats:italic>Staphylococcus aureus</jats:italic> that allows the deletion, mutation, or insertion of genes without leaving markers or other genetic scars. In addition, a protocol for rapid <jats:italic>in vitro</jats:italic> plasmid methylation and transformation is presented. The combined methods allow the routine genetic manipulation of <jats:italic>H. halodurans</jats:italic> from initial transformation to the desired strain in 8 days. These methods improve <jats:italic>H. halodurans</jats:italic> as a model organism for the study of extremophiles.","PeriodicalId":12466,"journal":{"name":"Frontiers in Microbiology","volume":null,"pages":null},"PeriodicalIF":4.0000,"publicationDate":"2024-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Frontiers in Microbiology","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.3389/fmicb.2024.1465811","RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"MICROBIOLOGY","Score":null,"Total":0}
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.
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.
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