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Applied and Environmental Microbiology, August 2001, p. 3350-3357, Vol. 67, No. 8
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

Jun Sun,1 Ilse Smets,2 Kristel Bernaerts,2 Jan Van Impe,2 Jos Vanderleyden,1,* and Kathleen Marchal3

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 beta -glucuronidase activity was used to monitor cytN gene expression. However, as the beta -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.


Applied and Environmental Microbiology, August 2001, p. 3350-3357, Vol. 67, No. 8
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.8.3350-3357.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.