| Registro completo |
| Provedor de dados: |
ArchiMer
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| País: |
France
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| Título: |
Genetic Manipulations of the Hyperthermophilic Piezophilic Archaeon Thermococcus barophilus
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| Autores: |
Thiel, Axel
Michoud, Gregoire
Moalic, Yann
Flament, Didier
Jebbar, Mohamed
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| Data: |
2014-04
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| Ano: |
2014
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| Resumo: |
In this study, we developed a gene disruption system for Thermococcus barophilus using simvastatin for positive selection and 5-fluoroorotic acid (5-FOA) for negative selection or counterselection to obtain markerless deletion mutants using single-and double-crossover events. Disruption plasmids carrying flanking regions of each targeted gene were constructed and introduced by transformation into wild-type T. barophilus MP cells. Initially, a pyrF deletion mutant was obtained as a starting point for the construction of further markerless mutants. A deletion of the hisB gene was also constructed in the UBOCC-3256 (Delta pyrF) background, generating a strain (UBOCC-3260) that was auxotrophic for histidine. A functional pyrF or hisB allele from T. barophilus was inserted into the chromosome of UBOCC-3256 (Delta pyrF) or UBOCC-3260 (Delta pyrF Delta hisB), allowing homologous complementation of these mutants. The piezophilic genetic tools developed in this study provide a way to construct strains with multiple genetic backgrounds that will allow further genetic studies for hyperthermophilic piezophilic archaea.
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| Tipo: |
Text
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| Idioma: |
Inglês
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| Identificador: |
http://archimer.ifremer.fr/doc/00187/29818/28263.pdf
DOI:10.1128/AEM.00084-14
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| Editor: |
Amer Soc Microbiology
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| Relação: |
http://archimer.ifremer.fr/doc/00187/29818/
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| Formato: |
application/pdf
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| Fonte: |
Applied And Environmental Microbiology (0099-2240) (Amer Soc Microbiology), 2014-04 , Vol. 80 , N. 7 , P. 2299-2306
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| Direitos: |
2014, American Society for Microbiology. All Rights Reserved.
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