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Applied and Environmental Microbiology, January 2000, p. 163-169, Vol. 66, No. 1
Environmental Molecular Biology Group,
Biosciences Division, Los Alamos National Laboratory, Los Alamos,
New Mexico 87545
Received 23 August 1999/Accepted 20 October 1999
The genetic systems of bacteria that have the ability to use
organic pollutants as carbon and energy sources can be adapted to
create bacterial biosensors for the detection of industrial pollution.
The creation of bacterial biosensors is hampered by a lack of
information about the genetic systems that control production of
bacterial enzymes that metabolize pollutants. We have attempted to
overcome this problem through modification of DmpR, a regulatory protein for the phenol degradation pathway of Pseudomonas
sp. strain CF600. The phenol detection capacity of DmpR was altered by
using mutagenic PCR targeted to the DmpR sensor domain. DmpR mutants
were identified that both increased sensitivity to the phenolic
effectors of wild-type DmpR and increased the range of molecules
detected. The phenol detection characteristics of seven DmpR mutants
were demonstrated through their ability to activate transcription of a
lacZ reporter gene. Effectors of the DmpR derivatives included phenol, 2-chlorophenol, 2,4-dichlorophenol,
4-chloro-3-methylphenol, 2,4-dimethylphenol, 2-nitrophenol, and
4-nitrophenol.
0099-2240/0/$04.00+0
Generation of Novel Bacterial Regulatory Proteins
That Detect Priority Pollutant Phenols
*
Corresponding author. Mailing address: Environmental
Molecular Biology Group, M888, Biosciences Division, Los Alamos
National Laboratory, Los Alamos, NM 87545. Phone: (505) 665-4800. Fax: (505) 665-3024. E-mail: kuske{at}lanl.gov.
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