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Title: Detection of phenols using engineered bacteria

Abstract

Detection of phenols using engineered bacteria. A biosensor can be created by placing a reporter gene under control of an inducible promoter. The reporter gene produces a signal when a cognate transcriptional activator senses the inducing chemical. Creation of bacterial biosensors is currently restricted by limited knowledge of the genetic systems of bacteria that catabolize xenobiotics. By using mutagenic PCR to change the chemical specificity of the Pseudomonas species CF600 DmpR protein, the potential for engineering novel biosensors for detection of phenols has been demonstrated. DmpR, a well-characterized transcriptional activator of the P. CF600's dmp operon mediates growth on simple phenols. Transcription from Po, the promoter heading the dmp operon, is activated when the sensor domain of DmpR interacts with phenol and mono-substituted phenols. By altering the sensor domain of the DmpR, a group of DmpR derivatives that activate transcription of a Po-lacZ fusion in response to eight of the EPA's eleven priority pollutant phenols has been created. The assays and the sensor domain mutations that alter the chemical specificity of DmpR is described.

Inventors:
; ;
Issue Date:
Research Org.:
Los Alamos National Lab. (LANL), Los Alamos, NM (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1174978
Patent Number(s):
6773918
Application Number:
09/520,538
Assignee:
University Of California, The Regents Of
Patent Classifications (CPCs):
C - CHEMISTRY C12 - BIOCHEMISTRY C12Q - MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
DOE Contract Number:  
W-7405-ENG-36
Resource Type:
Patent
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Wise, Arlene A., Kuske, Cheryl R., and Terwilliger, Thomas C. Detection of phenols using engineered bacteria. United States: N. p., 2004. Web.
Wise, Arlene A., Kuske, Cheryl R., & Terwilliger, Thomas C. Detection of phenols using engineered bacteria. United States.
Wise, Arlene A., Kuske, Cheryl R., and Terwilliger, Thomas C. Tue . "Detection of phenols using engineered bacteria". United States. https://www.osti.gov/servlets/purl/1174978.
@article{osti_1174978,
title = {Detection of phenols using engineered bacteria},
author = {Wise, Arlene A. and Kuske, Cheryl R. and Terwilliger, Thomas C.},
abstractNote = {Detection of phenols using engineered bacteria. A biosensor can be created by placing a reporter gene under control of an inducible promoter. The reporter gene produces a signal when a cognate transcriptional activator senses the inducing chemical. Creation of bacterial biosensors is currently restricted by limited knowledge of the genetic systems of bacteria that catabolize xenobiotics. By using mutagenic PCR to change the chemical specificity of the Pseudomonas species CF600 DmpR protein, the potential for engineering novel biosensors for detection of phenols has been demonstrated. DmpR, a well-characterized transcriptional activator of the P. CF600's dmp operon mediates growth on simple phenols. Transcription from Po, the promoter heading the dmp operon, is activated when the sensor domain of DmpR interacts with phenol and mono-substituted phenols. By altering the sensor domain of the DmpR, a group of DmpR derivatives that activate transcription of a Po-lacZ fusion in response to eight of the EPA's eleven priority pollutant phenols has been created. The assays and the sensor domain mutations that alter the chemical specificity of DmpR is described.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {2004},
month = {8}
}

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Works referenced in this record:

High efficiency transformation of E.coli by high voltage electroporation
journal, July 1988


Development and Testing of a Bacterial Biosensor for Toluene-Based Environmental Contaminants
journal, March 1998


Rapid evolution of a protein in vitro by DNA shuffling
journal, August 1994


Improved single and multicopy lac-based cloning vectors for protein and operon fusions
journal, January 1987