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Title: | An improved genome editing system for Sphingomonadaceae |
Other Titles: | An improved genome editing system for <i>Sphingomonadaceae</i> |
Authors: | García-Romero, I de Dios, R Reyes-Ramírez, F |
Keywords: | bioremediation;genetic tools;genome editing;mutation;Sphingomonadaceae;sphingomonads |
Issue Date: | 13-May-2024 |
Publisher: | Microbiology Society |
Citation: | García-Romero, I., de Dios, R. and Reyes-Ramírez, F. (2024) 'An improved genome editing system for Sphingomonadaceae', Access Microbiology, 6 (5), pp. 1 - 12. doi: 10.1099/acmi.0.000755.v3. |
Abstract: | The sphingomonads encompass a diverse group of bacteria within the family Sphingomonadaceae, with the presence of sphingolipids on their cell surface instead of lipopolysaccharide as their main common feature. They are particularly interesting for bioremediation purposes due to their ability to degrade or metabolise a variety of recalcitrant organic pollutants. However, research and development on their full bioremediation potential has been hampered because of the limited number of tools available to investigate and modify their genome. Here, we present a markerless genome editing method for Sphingopyxis granuli TFA, which can be further optimised for other sphingomonads. This procedure is based on a double recombination triggered by a DNA double-strand break in the chromosome. The strength of this protocol lies in forcing the second recombination rather than favouring it by pressing a counterselection marker, thus avoiding laborious restreaking or passaging screenings. Additionally, we introduce a modification with respect to the original protocol to increase the efficiency of the screening after the first recombination event. We show this procedure step by step and compare our modified method with respect to the original one by deleting ecfG2, the master regulator of the general stress response in S. granuli TFA. This adds to the genetic tool repertoire that can be applied to sphingomonads and stands as an efficient option for fast genome editing of this bacterial group. |
Description: | Data Summary:
All data and protocols used or generated through this work have been provided within this article or in the associated supplementary files. Two supplementary figures and two supplementary tables are available with the online version of this article. |
URI: | https://bura.brunel.ac.uk/handle/2438/28681 |
DOI: | https://doi.org/10.1099/acmi.0.000755.v3 |
Other Identifiers: | ORCiD: Inmaculada García-Romero https://orcid.org/0000-0002-8939-3271 ORCiD: Rubén de Dios https://orcid.org/0000-0001-6704-9149 ORCiD: Francisca Reyes-Ramírez https://orcid.org/0000-0002-5120-8931 |
Appears in Collections: | Dept of Life Sciences Research Papers |
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FullText.pdf | Copyright © 2024 The Authors. This is an open-access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/). | 1.08 MB | Adobe PDF | View/Open |
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