Benzoic Acid-Inducible Gene Expression in Mycobacteria
Conditional expression is a powerful tool to investigate the role of bacterial genes. Here, we adapt the Pseudomonas putida-derived positively regulated XylS/Pm expression system to control inducible gene expression in Mycobacterium smegmatis and Mycobacterium tuberculosis, the causative agent of hu...
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| Veröffentlicht in: | PloS one 2015-09, Vol.10 (9), p.e0134544-e0134544 |
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| creator | Dragset, Marte S Barczak, Amy K Kannan, Nisha Mærk, Mali Flo, Trude H Valla, Svein Rubin, Eric J Steigedal, Magnus |
| description | Conditional expression is a powerful tool to investigate the role of bacterial genes. Here, we adapt the Pseudomonas putida-derived positively regulated XylS/Pm expression system to control inducible gene expression in Mycobacterium smegmatis and Mycobacterium tuberculosis, the causative agent of human tuberculosis. By making simple changes to a Gram-negative broad-host-range XylS/Pm-regulated gene expression vector, we prove that it is possible to adapt this well-studied expression system to non-Gram-negative species. With the benzoic acid-derived inducer m-toluate, we achieve a robust, time- and dose-dependent reversible induction of Pm-mediated expression in mycobacteria, with low background expression levels. XylS/Pm is thus an important addition to existing mycobacterial expression tools, especially when low basal expression is of particular importance. |
| doi_str_mv | 10.1371/journal.pone.0134544 |
| format | Article |
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Here, we adapt the Pseudomonas putida-derived positively regulated XylS/Pm expression system to control inducible gene expression in Mycobacterium smegmatis and Mycobacterium tuberculosis, the causative agent of human tuberculosis. By making simple changes to a Gram-negative broad-host-range XylS/Pm-regulated gene expression vector, we prove that it is possible to adapt this well-studied expression system to non-Gram-negative species. With the benzoic acid-derived inducer m-toluate, we achieve a robust, time- and dose-dependent reversible induction of Pm-mediated expression in mycobacteria, with low background expression levels. XylS/Pm is thus an important addition to existing mycobacterial expression tools, especially when low basal expression is of particular importance.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0134544</identifier><identifier>PMID: 26348349</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Acids ; Bacteria ; Bacterial Proteins - biosynthesis ; Bacterial Proteins - genetics ; Benzoic acid ; Benzoic Acid - pharmacology ; Binding sites ; Cancer ; Drug dosages ; Drug resistance ; E coli ; Escherichia coli ; Gene expression ; Gene Expression Regulation, Bacterial ; Genetic aspects ; Genetic Vectors ; Gram-positive bacteria ; Humans ; Immunology ; Infectious diseases ; Inflammation ; Medical research ; Medicine ; Mycobacteria ; Mycobacterium smegmatis ; Mycobacterium smegmatis - genetics ; Mycobacterium smegmatis - pathogenicity ; Mycobacterium tuberculosis ; Mycobacterium tuberculosis - genetics ; Mycobacterium tuberculosis - pathogenicity ; Physiological aspects ; Promoter Regions, Genetic ; Pseudomonas putida ; Pseudomonas putida - genetics ; Public health ; Science ; Trans-Activators - biosynthesis ; Trans-Activators - genetics ; Tuberculosis ; Tuberculosis - genetics ; Tuberculosis - microbiology</subject><ispartof>PloS one, 2015-09, Vol.10 (9), p.e0134544-e0134544</ispartof><rights>COPYRIGHT 2015 Public Library of Science</rights><rights>2015 Dragset et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2015 Dragset et al 2015 Dragset et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c692t-9f0e6e3432c7e0d345d6d01eefecd65a8fd3691fba992b797bac4d96a59e87173</citedby><cites>FETCH-LOGICAL-c692t-9f0e6e3432c7e0d345d6d01eefecd65a8fd3691fba992b797bac4d96a59e87173</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4562662/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4562662/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,724,777,781,861,882,2096,2915,23847,27905,27906,53772,53774,79349,79350</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/26348349$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Kremer, Laurent</contributor><creatorcontrib>Dragset, Marte S</creatorcontrib><creatorcontrib>Barczak, Amy K</creatorcontrib><creatorcontrib>Kannan, Nisha</creatorcontrib><creatorcontrib>Mærk, Mali</creatorcontrib><creatorcontrib>Flo, Trude H</creatorcontrib><creatorcontrib>Valla, Svein</creatorcontrib><creatorcontrib>Rubin, Eric J</creatorcontrib><creatorcontrib>Steigedal, Magnus</creatorcontrib><title>Benzoic Acid-Inducible Gene Expression in Mycobacteria</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>Conditional expression is a powerful tool to investigate the role of bacterial genes. Here, we adapt the Pseudomonas putida-derived positively regulated XylS/Pm expression system to control inducible gene expression in Mycobacterium smegmatis and Mycobacterium tuberculosis, the causative agent of human tuberculosis. By making simple changes to a Gram-negative broad-host-range XylS/Pm-regulated gene expression vector, we prove that it is possible to adapt this well-studied expression system to non-Gram-negative species. With the benzoic acid-derived inducer m-toluate, we achieve a robust, time- and dose-dependent reversible induction of Pm-mediated expression in mycobacteria, with low background expression levels. XylS/Pm is thus an important addition to existing mycobacterial expression tools, especially when low basal expression is of particular importance.</description><subject>Acids</subject><subject>Bacteria</subject><subject>Bacterial Proteins - biosynthesis</subject><subject>Bacterial Proteins - genetics</subject><subject>Benzoic acid</subject><subject>Benzoic Acid - pharmacology</subject><subject>Binding sites</subject><subject>Cancer</subject><subject>Drug dosages</subject><subject>Drug resistance</subject><subject>E coli</subject><subject>Escherichia coli</subject><subject>Gene expression</subject><subject>Gene Expression Regulation, Bacterial</subject><subject>Genetic aspects</subject><subject>Genetic Vectors</subject><subject>Gram-positive bacteria</subject><subject>Humans</subject><subject>Immunology</subject><subject>Infectious diseases</subject><subject>Inflammation</subject><subject>Medical research</subject><subject>Medicine</subject><subject>Mycobacteria</subject><subject>Mycobacterium smegmatis</subject><subject>Mycobacterium smegmatis - 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| subjects | Acids Bacteria Bacterial Proteins - biosynthesis Bacterial Proteins - genetics Benzoic acid Benzoic Acid - pharmacology Binding sites Cancer Drug dosages Drug resistance E coli Escherichia coli Gene expression Gene Expression Regulation, Bacterial Genetic aspects Genetic Vectors Gram-positive bacteria Humans Immunology Infectious diseases Inflammation Medical research Medicine Mycobacteria Mycobacterium smegmatis Mycobacterium smegmatis - genetics Mycobacterium smegmatis - pathogenicity Mycobacterium tuberculosis Mycobacterium tuberculosis - genetics Mycobacterium tuberculosis - pathogenicity Physiological aspects Promoter Regions, Genetic Pseudomonas putida Pseudomonas putida - genetics Public health Science Trans-Activators - biosynthesis Trans-Activators - genetics Tuberculosis Tuberculosis - genetics Tuberculosis - microbiology |
| title | Benzoic Acid-Inducible Gene Expression in Mycobacteria |
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