PMID: 9435063Jan 22, 1998Paper

The sequence of spacers between the consensus sequences modulates the strength of prokaryotic promoters

Applied and Environmental Microbiology
P R Jensen, K Hammer

Abstract

We constructed a library of synthetic promoters for Lactococcus lactis in which the known consensus sequences were kept constant while the sequences of the separating spacers were randomized. The library consists of 38 promoters which differ in strength from 0.3 up to more than 2,000 relative units, the latter among the strongest promoters known for this organism. The ranking of the promoter activities was somewhat different when assayed in Escherichia coli, but the promoters are efficient for modulating gene expression in this bacterium as well. DNA sequencing revealed that the weaker promoters (which had activities below 5 relative units) all had changes either in the consensus sequences or in the length of the spacer between the -35 and -10 sequences. The promoters in which those features were conserved had activities from 5 to 2,050 U, which shows that by randomizing the spacers, at least a 400-fold change in activity can be obtained. Interestingly, the entire range of promoter activities is covered in small steps of activity increase, which makes these promoters very suitable for quantitative physiological studies and for fine-tuning of gene expression in industrial bioreactors and cell factories.

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Citations

Apr 27, 2002·Applied and Environmental Microbiology·Christian Solem, Peter Ruhdal Jensen
Oct 31, 2002·Applied and Environmental Microbiology·Martin B PedersenDan Nilsson
Aug 27, 2005·Proceedings of the National Academy of Sciences of the United States of America·Hal AlperGregory Stephanopoulos
Nov 5, 2005·Applied and Environmental Microbiology·Sung Kuk Lee, Jay D Keasling
Feb 10, 2009·Applied and Environmental Microbiology·Ingrid BakkeSvein Valla
Mar 2, 2010·Chembiochem : a European Journal of Chemical Biology·Claudia Ruth, Anton Glieder
Mar 20, 2010·The Journal of Biological Chemistry·Anders K HolmGoutham N Vemuri
Feb 13, 2010·Journal of Biomedicine & Biotechnology·Eric Young, Hal Alper
Apr 2, 2011·BMC Molecular Biology·Anthony R Poteete
Jul 9, 2011·Nucleic Acids Research·Andrew H Babiskin, Christina D Smolke
Jun 5, 2013·Biotechnology and Bioengineering·Bradley Niesner, Narendra Maheshri
Sep 29, 2015·Molecular Microbiology·Maksym Bobrovskyy, Carin K Vanderpool
Jun 26, 2002·Journal of Bacteriology·Brian J KoebmannPeter R Jensen
Jun 6, 2003·Applied and Environmental Microbiology·David A SchofieldMichael G Schmidt
Apr 7, 2005·Applied and Environmental Microbiology·Isabelle Meynial-SallesPhilippe Soucaille
Nov 18, 2004·Journal of Bacteriology·Anja H SchiemannPaul W O'Toole
Sep 28, 2005·Biotechnology and Bioengineering·Andrea ZanzottoKlavs F Jensen
Mar 5, 2005·Applied and Environmental Microbiology·Geneviève LamotheSylvain Moineau
May 23, 2006·Pharmaceutical Research·Ethlinn V B van GaalEnrico Mastrobattista
Feb 18, 2011·Plant Molecular Biology·Rajesh MehrotraSandhya Mehrotra
Nov 1, 2008·Biotechnology and Bioengineering·Yuansheng YangMiranda G S Yap
Sep 19, 2012·Proceedings of the National Academy of Sciences of the United States of America·Jennifer B RiceCarin K Vanderpool
Oct 28, 2010·Lab on a Chip·Sahar MelamedShimshon Belkin
Mar 9, 2013·Nucleic Acids Research·Shawn P Finney-Manchester, Narendra Maheshri
Mar 3, 2011·Molecular Systems Biology·Andrew H Babiskin, Christina D Smolke
Nov 28, 2012·Biochemical Society Transactions·Katrin Richter, Johannes Gescher

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