Data/hora: 06/06/2024 07:26:03

Provedor de dados: 	5	
País: 	France	
Título: 	Genetic Manipulations of the Hyperthermophilic Piezophilic Archaeon Thermococcus barophilus	
Autores: 	Thiel, Axel; Michoud, Gregoire; Moalic, Yann; Flament, Didier; Jebbar, Mohamed.	
Data: 	2014-04	
Ano: 	2014	
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.	
Tipo: 	Text	
Idioma: 	Inglês	
Identificador: 	http://archimer.ifremer.fr/doc/00187/29818/28263.pdf

DOI:10.1128/AEM.00084-14	
Editor: 	Amer Soc Microbiology	
Relação: 	http://archimer.ifremer.fr/doc/00187/29818/	
Formato: 	application/pdf	
Fonte: 	Applied And Environmental Microbiology (0099-2240) (Amer Soc Microbiology), 2014-04 , Vol. 80 , N. 7 , P. 2299-2306	
Fonte: 	2014, American Society for Microbiology. All Rights Reserved.