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Applied and Environmental Microbiology, November 1998, p. 4321-4327, Vol. 64, No. 11
Department of Physiology and Metabolism,
Received 23 February 1998/Accepted 11 August 1998
An operon containing the genes purD and
purE and part of the purK gene was cloned from
the facultative anaerobic gram-positive bacterium Lactococcus
lactis by complementation of the purD mutation in
Escherichia coli SØ609. The genes encode enzymes in the de novo pathway of purine nucleotides. The expression of the genes was
regulated approximately 35-fold at the transcription level by the
availability of purines in the growth medium. Deletion analysis of the
nucleotide region upstream of purD indicated that a region
of 145 bp is enough to give regulated expression of the reporter
lacLM genes, which encode
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Cloning and Expression of the Lactococcus
lactis purDEK Genes, Required for Growth in Milk
-galactosidase. Deletion of a
region 79 bp upstream of the transcription start point reduced the
promoter activity 33-fold when incubated in a purine-free medium and to
values below the detection limit when incubated in a purine-containing
medium. No secondary transcription start points were mapped in or close
to this region, indicating that a putative activator site and not a
promoter was deleted or partly destroyed.
*
Corresponding author. Mailing address: Department of
Physiology and Metabolism, Chr. Hansen A/S, Bøge Allé 10-12, DK-2970 Hørsholm, Denmark. Phone: 45 45 74 74 74. Fax: 45 45 74 89 94. E-mail: dn.dk{at}chr-hansen.com.
Applied and Environmental Microbiology, November 1998, p. 4321-4327, Vol. 64, No. 11
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
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