| [PMID]: | 28257493 |
| [Au] Autor: | Gebhardt MJ; Jacobson RK; Shuman HA |
| [Ad] Endereço: | Department of Microbiology, University of Chicago, Chicago, Illinois, United States of America. |
| [Ti] Título: | Seeing red; the development of pON.mCherry, a broad-host range constitutive expression plasmid for Gram-negative bacteria. |
| [So] Source: | PLoS One;12(3):e0173116, 2017. |
| [Is] ISSN: | 1932-6203 |
| [Cp] País de publicação: | United States |
| [La] Idioma: | eng |
| [Ab] Resumo: | The development of plasmid-mediated gene expression control in bacteria revolutionized the field of bacteriology. Many of these expression control systems rely on the addition of small molecules, generally metabolites or non-metabolized analogs thereof, to the growth medium to induce expression of the genes of interest. The paradigmatic example of an expression control system is the lac system from Escherichia coli, which typically relies on the Ptac promoter and the Lac repressor, LacI. In many cases, however, constitutive gene expression is desired, and other experimental approaches require the coordinated control of multiple genes. While multiple systems have been developed for use in E. coli and its close relatives, the utility and/or functionality of these tools does not always translate to other species. For example, for the Gram-negative pathogen, Legionella pneumophila, a causative agent of Legionnaires' Disease, the aforementioned Ptac system represents the only well-established expression control system. In order to enhance the tools available to study bacterial gene expression in L. pneumophila, we developed a plasmid, pON.mCherry, which confers constitutive gene expression from a mutagenized LacI binding site. We demonstrate that pON.mCherry neither interferes with other plasmids harboring an intact LacI-Ptac expression system nor alters the growth of Legionella species during intracellular growth. Furthermore, the broad-host range plasmid backbone of pON.mCherry allows constitutive gene expression in a wide variety of Gram-negative bacterial species, making pON.mCherry a useful tool for the greater research community. |
| [Mh] Termos MeSH primário: |
Escherichia coli/genética
Regulação Bacteriana da Expressão Gênica
Legionella pneumophila/genética
Proteínas Luminescentes/genética
Plasmídeos/química
|
| [Mh] Termos MeSH secundário: |
Escherichia coli/metabolismo
Escherichia coli/ultraestrutura
Engenharia Genética
Repressores Lac/genética
Repressores Lac/metabolismo
Legionella pneumophila/metabolismo
Legionella pneumophila/ultraestrutura
Proteínas Luminescentes/metabolismo
Biologia Molecular/métodos
Mutação
Plasmídeos/metabolismo
Regiões Promotoras Genéticas
|
| [Pt] Tipo de publicação: | JOURNAL ARTICLE |
[Nm] Nome de substância:
| 0 (Lac Repressors); 0 (Luminescent Proteins); 0 (red fluorescent protein) |
| [Em] Mês de entrada: | 1708 |
| [Cu] Atualização por classe: | 170828 |
[Lr] Data última revisão:
| 170828 |
| [Sb] Subgrupo de revista: | IM |
| [Da] Data de entrada para processamento: | 170304 |
| [St] Status: | MEDLINE |
| [do] DOI: | 10.1371/journal.pone.0173116 |
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