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PMID: 3525513 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Vector for regulated expression of cloned genes in a wide range of gram-negative bacteria.

Journal of bacteriology ·Vol. 167 ·No. 2 ·1986-08-00 ·Pages 447-54

Mermod N, Ramos JL, Lehrbach PR, Timmis KN

Abstract

A pKT231-based broad-host-range plasmid vector was constructed which enabled regulation of expression of cloned genes in a wide range of gram-negative bacteria. This vector, pNM185, contained upstream of its EcoRI, SstI, and SstII cloning sites the positively activated pm twin promoters of the TOL plasmid and xylS, the gene of the positive regulator of these promoters. Expression of cloned genes was induced with micromolar quantities of benzoate or m-toluate, the inexpensive coinducers of the pm promoters. Expression of a test gene, xylE, which specifies catechol 2,3-dioxygenase, cloned in this vector was tested in representative strains of a variety of gram-negative bacteria. Regulated expression of xylE was observed in most strains examined, and induced levels of enzyme representing up to 5% of total cellular protein and ratios of induced:noninduced levels of enzyme up to a factor of 600 were observed. The level of xylE gene expression in different bacteria tended to be correlated with their phylogenetic distance from Pseudomonas putida.

MeSH Terms
Drug Resistance, Microbial Escherichia coli/genetics Gene Expression Regulation Genes, Bacterial Genetic Vectors Gram-Negative Bacteria/genetics Plasmids Promoter Regions, Genetic Pseudomonas/genetics Species Specificity Streptomycin/pharmacology
Chemicals
Streptomycin
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Mermod N
Ramos J L
Lehrbach P R
Timmis K N
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36 references, click to expand
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.167.2.447-454.1986
Published
1986-08-00
Pages
447-54
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC212908
Subset
IM
External Links
PubMed source DOI: 10.1128/jb.167.2.447-454.1986 PMC Full Text View journal details
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