PMID: 8425528Jan 15, 1993Paper

The use of lac-type promoters in control analysis

European Journal of Biochemistry
P R JensenO Michelsen

Abstract

For control analysis, it is necessary to modulate the activity of an enzyme around its normal level and measure the changes in steady-state fluxes or concentrations. We describe an improved method for effecting the modulation, as elaborated for Escherichia coli. The chromosomal gene, encoding the enzyme of interest, is put under the control of a lacUV5 or a tacI promoter. The alternative use of the two promoters leads to an expression range which should make it suitable for the use in control analysis of many enzymes. The lacUV5 promoter should be used when the wild-type expression level is low, the tacI promoter when the latter is high. The endogenous lac operon is placed under the control of a second copy of the lacUV5 promoter and a lacY7am mutation (eliminating lactose permease, the transport system for the inducer isopropyl-thio-beta-D- galactoside) is introduced. The method was demonstrated experimentally by constructing E. coli strains, in which the chromosomal atp operon is transcribed from the lacUV5 and the tacI promoter. We measured the concentration of the c subunit of H(+)-ATPase, and found that the expression of this enzyme could be modulated between non-detectable levels and up to five times the wild-type level. ...Continue Reading

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Citations

Dec 1, 1995·Journal of Bioenergetics and Biomembranes·P R JensenH V Westerhoff
Nov 15, 2003·Journal of Theoretical Biology·Mark A PeletierBoris N Kholodenko
Feb 7, 2001·Mutation Research·L BoeP R Jensen
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Sep 17, 2002·European Journal of Biochemistry·Alberto de la FuentePedro Mendes
Mar 23, 2007·Proceedings of the National Academy of Sciences of the United States of America·Thomas KuhlmanTerence Hwa
Jun 27, 2013·Proceedings of the National Academy of Sciences of the United States of America·Joseph P TorellaPamela A Silver
Sep 1, 1993·Proceedings of the National Academy of Sciences of the United States of America·P R JensenH V Westerhoff
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Apr 27, 2002·Applied and Environmental Microbiology·Christian Solem, Peter Ruhdal Jensen
Nov 5, 2005·Applied and Environmental Microbiology·Sung Kuk Lee, Jay D Keasling
Jun 26, 2002·Journal of Bacteriology·Brian J KoebmannPeter R Jensen
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Jan 25, 2014·Applied Microbiology and Biotechnology·Jakob Vang RytterPeter Ruhdal Jensen
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Jun 18, 2005·Current Opinion in Biotechnology·Ivan MijakovicPeter Ruhdal Jensen
Jul 15, 2015·FEMS Microbiology Letters·Duolong ZhuMingqiang Qiao
Oct 4, 2016·Frontiers in Cellular and Infection Microbiology·Hannes Breddermann, Karin Schnetz
Nov 15, 2016·Nature Chemical Biology·Dan I PiranerMikhail G Shapiro
Dec 22, 2016·Applied Microbiology and Biotechnology·Lukas MarschallChristoph Herwig
Aug 1, 2015·Molecular BioSystems·Huy TranAndre S Ribeiro
May 14, 2016·Applied Microbiology and Biotechnology·Lukas MarschallChristoph Herwig
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Mar 29, 1993·FEBS Letters·B N Kholodenko, H V Westerhoff
Jan 22, 1998·Applied and Environmental Microbiology·P R Jensen, K Hammer
Nov 13, 1998·Journal of Bacteriology·F C BoogerdP R Jensen

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