FN Archimer Export Format
PT J
TI Genetic Manipulations of the Hyperthermophilic Piezophilic Archaeon Thermococcus barophilus
BT 
AF THIEL, Axel
   MICHOUD, Gregoire
   MOALIC, Yann
   FLAMENT, Didier
   JEBBAR, Mohamed
AS 1:1,2,3;2:1,2,3;3:1,2,3;4:1,2,3;5:1,2,3;
FF 1:PDG-REM-EEP-LMEE;2:;3:PDG-REM-EEP-LEP;4:PDG-REM-EEP-LMEE;5:;
C1 Univ Bretagne Occidentale, UMR 6197, Lab Microbiol Environm Extremes, Inst Univ Europeen Mer, Plouzane, France.
   CNRS, UMR 6197, Lab Microbiol Environm Extremes, Inst Univ Europeen Mer, Plouzane, France.
   IFREMER, UMR 6197, Lab Microbiol Environm Extremes, Plouzane, France.
C2 UBO, FRANCE
   CNRS, FRANCE
   IFREMER, FRANCE
SI BREST
SE PDG-REM-EEP-LMEE
   PDG-REM-EEP-LEP
IN WOS Ifremer jusqu'en 2018
   copubli-france
   copubli-univ-france
IF 3.668
TC 22
UR https://archimer.ifremer.fr/doc/00187/29818/28263.pdf
LA English
DT Article
AB 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.
PY 2014
PD APR
SO Applied And Environmental Microbiology
SN 0099-2240
PU Amer Soc Microbiology
VL 80
IS 7
UT 000332840700029
BP 2299
EP 2306
DI 10.1128/AEM.00084-14
ID 29818
ER

EF