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Applied and Environmental Microbiology, August 2001, p. 3350-3357, Vol. 67, No. 8
Centre of Microbial and Plant
Genetics,1 BioTeC-Bioprocess Technology
and Control,2 and SISTA, Department
of Electrical Engineering,3 Katholieke
Universiteit Leuven, B-3001 Heverlee, Belgium
Received 21 December 2000/Accepted 2 May 2001
An Azospirillum brasilense Sp7 strain containing a
plasmid-borne translational cytN-gusA fusion was grown in a
continuous culture to quantitatively evaluate the influence of
extracellular signals (such as O2) on expression of the
cytNOQP operon. The dissolved oxygen concentration was
shifted at regular time intervals before the steady state was reached.
The measured
0099-2240/01/$04.00+0 DOI: 10.1128/AEM.67.8.3350-3357.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Quantitative Analysis of Bacterial Gene Expression
by Using the gusA Reporter Gene System
-glucuronidase activity was used to monitor
cytN gene expression. However, as the
-glucuronidase
activity in the experimental setup not only depended on altered
transcription of the hybrid gene when the signal was varied but was
also influenced by cellular accumulation, degradation, and dilution of
the hybrid fusion protein, a mathematical method was developed to
describe the intrinsic properties of the dynamic bioprocess. After
identification and validation of the mathematical model, the apparent
specific rate of expression of the fusion, which was independent of the
experimental setup, could be deduced from the model and used to
quantify gene expression regulated by extracellular environmental
signals. In principle, this approach can be generalized to assess the
effects of external signals on bacterial gene expression.
*
Corresponding author. Mailing address: Centre of
Microbial and Plant Genetics, K. U. Leuven, Kasteelpark Arenberg
20, B-3001 Heverlee, Belgium. Phone: 0032-16-321631. Fax:
0032-16-321966. E-mail:
jozef.vanderleyden{at}agr.kuleuven.ac.be.
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