Nucleic Acids Research, Vol 25, Issue 6 1203-1210, Copyright © 1997 by Oxford University Press
R Lutz and H Bujard
Based on parameters governing promoter activity and using regulatory
elements of the lac, ara and tet operon transcription control sequences
were composed which permit the regulation in Escherichia coli of several
gene activities independently and quantitatively. The novel promoter
PLtetO-1 allows the regulation of gene expression over an up to 5000-fold
range with anhydrotetracycline (aTc) whereas with IPTG and arabinose the
activity of Plac/ara-1 may be controlled 1800-fold. Escherichia coli host
strains which produce defined amounts of the regulatory proteins, Lac and
Tet repressor as well as AraC from chromosomally located expression units
provide highly reproducible in vivo conditions. Controlling the expression
of the genes encoding luciferase, the low abundance E.coli protein DnaJ and
restriction endonuclease Cfr9I not only demonstrates that high levels of
expression can be achieved but also suggests that under conditions of
optimal repression only around one mRNA every 3rd generation is produced.
This potential of quantitative control will open up new approaches in the
study of gene function in vivo, in particular with low abundance regulatory
gene products. The system will also provide new opportunities for the
controlled expression of heterologous genes.
ARTICLES
Independent and tight regulation of transcriptional units in Escherichia coli via the LacR/O, the TetR/O and AraC/I1-I2 regulatory elements
ZMBH Zentrum fur Molekulare Biologie der Universitat Heidelberg, Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany.
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