Improved methods for genetic manipulation of the alkaliphile Halalkalibacterium halodurans .
| Autor: | Wencker FDR; Howard Hughes Medical Institute, Yale University, New Haven, CT, United States., Lyon SE; Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT, United States., Breaker RR; Howard Hughes Medical Institute, Yale University, New Haven, CT, United States.; Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT, United States.; Department of Molecular, Cellular and Developmental Biology, Yale University, New Haven, CT, United States. |
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| Jazyk: | angličtina |
| Zdroj: | Frontiers in microbiology [Front Microbiol] 2024 Sep 18; Vol. 15, pp. 1465811. Date of Electronic Publication: 2024 Sep 18 (Print Publication: 2024). |
| DOI: | 10.3389/fmicb.2024.1465811 |
| Abstrakt: | 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. Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. (Copyright © 2024 Wencker, Lyon and Breaker.) |
| Databáze: | MEDLINE |
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