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Journal articles
Genetic manipulations of the hyperthermophilic piezophilic archaeon Thermococcus barophilus.
Abstract : 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 (ΔpyrF) 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 (ΔpyrF) or UBOCC-3260 (ΔpyrF ΔhisB), 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.
Cited literature [29 references]
https://hal.univ-brest.fr/hal-01019712
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Submitted on : Monday, July 7, 2014 - 1:51:40 PM
Last modification on : Monday, October 11, 2021 - 2:22:07 PM
Long-term archiving on: : Tuesday, October 7, 2014 - 4:52:04 PM
Contributor : Adminhal Univbrestbu Connect in order to contact the contributor
Submitted on : Monday, July 7, 2014 - 1:51:40 PM
Last modification on : Monday, October 11, 2021 - 2:22:07 PM
Long-term archiving on: : Tuesday, October 7, 2014 - 4:52:04 PM
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Appl._Environ._Microbiol.-2014...
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