scholarly journals High hydrostatic pressure adaptive strategies in an obligate piezophile Pyrococcus yayanosii

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Grégoire Michoud ◽  
Mohamed Jebbar
Keyword(s):  
Hydrostatic Pressure ◽  
High Hydrostatic Pressure ◽  
Adaptive Strategies ◽  
Pyrococcus Yayanosii

scholarly journals Induction of a Toxin-Antitoxin Gene Cassette under High Hydrostatic Pressure Enables Markerless Gene Disruption in the Hyperthermophilic Archaeon Pyrococcus yayanosii

2018 ◽  
Vol 85 (4) ◽  
Author(s):  
Qinghao Song ◽  
Zhen Li ◽  
Rouke Chen ◽  
Xiaopan Ma ◽  
Xiang Xiao ◽  
...  
Keyword(s):  
Hydrostatic Pressure ◽  
High Temperatures ◽  
High Hydrostatic Pressure ◽  
Gene Disruption ◽  
Inducible Promoter ◽  
Constitutive Promoter ◽  
Gene Encoding ◽  
Content Type ◽  
Genetic Tools ◽  
Pyrococcus Yayanosii

ABSTRACT The discovery of hyperthermophiles has dramatically changed our understanding of the habitats in which life can thrive. However, the extreme high temperatures in which these organisms live have severely restricted the development of genetic tools. The archaeon Pyrococcus yayanosii A1 is a strictly anaerobic and piezophilic hyperthermophile that is an ideal model for studies of extreme environmental adaptation. In the present study, we identified a high hydrostatic pressure (HHP)-inducible promoter (Phhp) that controls target gene expression under HHP. We developed an HHP-inducible toxin-antitoxin cassette (HHP-TAC) containing (i) a counterselectable marker in which a gene encoding a putative toxin (virulence-associated protein C [PF0776 {VapC}]) controlled by the HHP-inducible promoter was used in conjunction with the gene encoding antitoxin PF0775 (VapB), which was fused to a constitutive promoter (PhmtB), and (ii) a positive marker with the 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase-encoding gene from P. furiosus controlled by the constitutive promoter Pgdh. The HHP-TAC was constructed to realize markerless gene disruption directly in P. yayanosii A1 in rich medium. The pop-out recombination step was performed using an HHP-inducible method. As proof, the PYCH_13690 gene, which encodes a 4-α-glucanotransferase, was successfully deleted from the strain P. yayanosii A1. The results showed that the capacity for starch hydrolysis in the Δ1369 mutant decreased dramatically compared to that in the wild-type strain. The inducible toxin-antitoxin system developed in this study greatly increases the genetic tools available for use in hyperthermophiles. IMPORTANCE Genetic manipulations in hyperthermophiles have been studied for over 20 years. However, the extremely high temperatures under which these organisms grow have limited the development of genetic tools. In this study, an HHP-inducible promoter was used to control the expression of a toxin. Compared to sugar-inducible and cold-shock-inducible promoters, the HHP-inducible promoter rarely has negative effects on the overall physiology and central metabolism of microorganisms, especially piezophilic hyperthermophiles. Previous studies have used auxotrophic strains as hosts, which may interfere with studies of adaptation and metabolism. Using an inducible toxin-antitoxin (TA) system as a counterselectable marker enables the generation of a markerless gene disruption strain without the use of auxotrophic mutants and counterselection with 5-fluoroorotic acid. TA systems are widely distributed in bacteria and archaea and can be used to overcome the limitations of high growth temperatures and dramatically extend the selectivity of genetic tools in hyperthermophiles.

Survival of mouse blastocysts after low-temperature preservation under high pressure

2004 ◽  
Vol 52 (4) ◽  
pp. 479-487 ◽  
Author(s):  
Cs. Pribenszky ◽  
M. Molnár ◽  
S. Cseh ◽  
L. Solti
Keyword(s):  
Phase Transition ◽  
Hydrostatic Pressure ◽  
Low Temperature ◽  
High Hydrostatic Pressure ◽  
Room Temperature ◽  
Freezing Point ◽  
Significant Loss ◽  
Embryo Cryopreservation ◽  
Subzero Temperatures ◽  
Survival Capacity

Cryoinjuries are almost inevitable during the freezing of embryos. The present study examines the possibility of using high hydrostatic pressure to reduce substantially the freezing point of the embryo-holding solution, in order to preserve embryos at subzero temperatures, thus avoiding all the disadvantages of freezing. The pressure of 210 MPa lowers the phase transition temperature of water to -21°C. According to the results of this study, embryos can survive in high hydrostatic pressure environment at room temperature; the time embryos spend under pressure without significant loss in their survival could be lengthened by gradual decompression. Pressurisation at 0°C significantly reduced the survival capacity of the embryos; gradual decompression had no beneficial effect on survival at that stage. Based on the findings, the use of the phenomena is not applicable in this form, since pressure and low temperature together proved to be lethal to the embryos in these experiments. The application of hydrostatic pressure in embryo cryopreservation requires more detailed research, although the experience gained in this study can be applied usefully in different circumstances.

Effect of High Hydrostatic Pressure on Endothelial Lipase Expression*

2010 ◽  
Vol 37 (6) ◽  
pp. 641-645 ◽  
Author(s):  
Can-Xin XU ◽  
Chun WANG ◽  
Bing-Yang ZHU ◽  
Zhi-Ping GAO ◽  
Di-Xian LUO ◽  
...  
Keyword(s):  
Hydrostatic Pressure ◽  
High Hydrostatic Pressure ◽  
Endothelial Lipase ◽  
Lipase Expression
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