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Applied and Environmental Microbiology, November 2005, p. 6856-6862, Vol. 71, No. 11
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.11.6856-6862.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

A Propionate-Inducible Expression System for Enteric Bacteria

Sung Kuk Lee and Jay D. Keasling*

Departments of Chemical Engineering and Bioengineering, University of California, and Synthetic Biology Department, Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720

Received 11 May 2005/ Accepted 26 June 2005

A series of new expression vectors (pPro) have been constructed for the regulated expression of genes in Escherichia coli. The pPro vectors contain the prpBCDE promoter (PprpB) responsible for expression of the propionate catabolic genes (prpBCDE) and prpR encoding the positive regulator of this promoter. The efficiency and regulatory properties of the prpR-PprpB system were measured by placing the gene encoding the green fluorescent protein (gfp) under the control of the inducible PprpB of E. coli. This system provides homogenous expression in individual cells, highly regulatable expression over a wide range of propionate concentrations, and strong expression (maximal 1,500-fold induction) at high propionate concentrations. Since the prpBCDE promoter has CAP-dependent activation, the prpR-PprpB system exhibited negligible basal expression by addition of glucose to the medium.

* Corresponding author. Mailing address: Berkeley Center for Synthetic Biology, 717 Potter St., Bldg. 977, Mail Code 3224, University of California, Berkeley, CA 94720-3224. Phone: (510) 495-2620. Fax: (510) 495-2630. E-mail: keasling{at}berkeley.edu.

Applied and Environmental Microbiology, November 2005, p. 6856-6862, Vol. 71, No. 11
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.11.6856-6862.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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