Development and use of genetic system to identify genes required for efficient low-temperature growth of Psychrobacter arcticus 273-4

Extremophiles : Life Under Extreme Conditions
Corien BakermansMichael F Thomashow

Abstract

We describe the development of genetic tools (electroporation, conjugation, vector for targeted gene replacement) for use in the psychrophile Psychrobacter arcticus 273-4 to test hypotheses about cold adaptation. Successful electroporation only occurred with nonstandard parameters, such as: electrocompetent cells freshly prepared from stationary-phase cultures, high field strengths (25 kV cm(-1)), long recovery times (16-24 h), and selection with low concentrations of antibiotics. Transformation frequencies were greatly affected by a methylation-dependent restriction barrier homologous to DpnI. The vector pJK100 (which was self-transmissible and contained a Pir-dependent R6K origin of replication) proved effective as a suicide plasmid that could be used to recombine mutations into the P. arcticus 273-4 genome. We used this vector for targeted replacement of dctT, the substrate-binding periplasmic subunit of a TRAP (tripartite ATP-independent periplasmic) transporter (which we have named dctTUF), as it was more highly expressed at cold temperatures. The replacement of dctT (with kan) decreased the rate of growth at low temperatures in mineral medium with glutamate, acetate, butyrate, and fumarate, but not with pyruvate suggestin...Continue Reading

References

Jan 1, 1977·Comparative Biochemistry and Physiology. B, Comparative Biochemistry·N H Marcus
Jan 1, 1979·Comparative Biochemistry and Physiology. B, Comparative Biochemistry·H Yamawaki, H Tsukuda
Apr 1, 1979·Proceedings of the National Academy of Sciences of the United States of America·D H Figurski, D R Helinski
Jun 1, 1988·Journal of Bacteriology·N M Calvin, P C Hanawalt
Dec 1, 1980·Proceedings of the National Academy of Sciences of the United States of America·G DittaD R Helinski
Mar 1, 1997·Letters in Applied Microbiology·C R Myers, J M Myers
Jul 2, 1998·Applied and Environmental Microbiology·S StrettonA E Goodman
Jul 6, 2000·Extremophiles : Life Under Extreme Conditions·Tatiana VishnivetskayaJames M Tiedje
Jul 29, 2000·Biochemical and Biophysical Research Communications·M L TutinoG Marino
Aug 29, 2001·FEMS Microbiology Reviews·David J Kelly, Gavin H Thomas
Jul 20, 2002·Photochemistry and Photobiology·Sharon VanounouItzhak Fishov
Nov 27, 2002·BioTechniques·Christopher J Marx, Mary E Lidstrom
Mar 29, 2003·Environmental Microbiology·Corien BakermansKenneth H Nealson
Feb 10, 2004·Applied and Environmental Microbiology·Michael V TyurinLee R Lynd
Apr 20, 2004·Applied Microbiology and Biotechnology·L BeneyP Gervais
Dec 21, 2004·Nucleic Acids Research·Nicola J MulderCathy H Wu
Dec 7, 2005·FEMS Microbiology Ecology·Monica A PonderJames M Tiedje
Jun 2, 2006·International Journal of Systematic and Evolutionary Microbiology·Corien BakermansJames M Tiedje
Nov 24, 2006·Extremophiles : Life Under Extreme Conditions·Corien BakermansMichael F Thomashow
Sep 1, 1993·World Journal of Microbiology & Biotechnology·R M Atlas

Citations

Jul 29, 2010·Environmental Technology·Rosa Margesin, Georges Feller
Apr 16, 2011·Canadian Journal of Microbiology·Selim CeylanDilek Kazan
Sep 10, 2013·Applied and Environmental Microbiology·Eric M LennemanBrett M Barney
May 12, 2012·Journal of Bioscience and Bioengineering·Selim CeylanDilek Kazan
Jun 30, 2010·FEMS Microbiology Reviews·Christopher MulliganGavin H Thomas
Jul 27, 2010·Trends in Microbiology·Marcus FischerGavin H Thomas
Jul 6, 2013·FEMS Microbiology Letters·Jiashi WeiBrett M Barney
Mar 12, 2011·Die Naturwissenschaften·Francesco Canganella, Juergen Wiegel
Feb 27, 2018·Frontiers in Cellular and Infection Microbiology·Leonardo T RosaDavid J Kelly

Related Concepts

Adaptation, Physiological
Cold Temperature
Genes, Bacterial
Shuttle Vectors
Membrane Fluidity
Episomes
Electroporation
Reverse Transcriptase Polymerase Chain Reaction
Psychrobacter
Acetate

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