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PMID: 16269719 Published · ppublish English Evaluation Study Journal Article Research Support, N.I.H., Extramural

A propionate-inducible expression system for enteric bacteria.

Applied and environmental microbiology ·Vol. 71 ·No. 11 ·2005-11-00 ·Pages 6856-62

Lee SK, Keasling JD

Abstract

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 (P(prpB)) 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-P(prpB) system were measured by placing the gene encoding the green fluorescent protein (gfp) under the control of the inducible P(prpB) 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-P(prpB) system exhibited negligible basal expression by addition of glucose to the medium.

MeSH Terms
Bacterial Proteins/genetics,metabolism Base Sequence Citrates/metabolism Escherichia coli/genetics,metabolism Escherichia coli Proteins/genetics,metabolism Gene Expression Regulation, Bacterial Genetic Vectors/genetics Green Fluorescent Proteins/genetics,metabolism Molecular Sequence Data Plasmids/genetics Promoter Regions, Genetic/genetics Propionates/metabolism
Chemicals
Bacterial Proteins Citrates Escherichia coli Proteins Propionates Green Fluorescent Proteins 2-methylcitric acid
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Lee Sung Kuk
Department of Chemical Engineering, University of California, Berkeley, CA 94720, USA.
Keasling Jay D
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Article Info
Journal
Applied and environmental microbiology
Abbr.
Appl Environ Microbiol
ISSN
0099-2240
DOI
10.1128/AEM.71.11.6856-6862.2005
Published
2005-11-00
Pages
6856-62
Language
English
Region
United States
NLM ID
7605801
PMCID
PMC1287719
Subset
IM
Grants
NIGMS NIH HHS · R01 GM070763 · United States
NIGMS NIH HHS · GM070763-01 · United States
External Links
PubMed source DOI: 10.1128/AEM.71.11.6856-6862.2005 PMC Full Text View journal details
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