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Assessment of tfdR and tfdS Promoter Activity of 2,4-Dichlorophenoxyacetic Acid

EPA Grant Number: MA916288
Title: Assessment of tfdR and tfdS Promoter Activity of 2,4-Dichlorophenoxyacetic Acid
Investigators: Davison, Michelle
Institution: California State University - Fresno
EPA Project Officer: Willett, Stephanie H.
Project Period: January 1, 2004 through December 31, 2006
Project Amount: $88,478
RFA: GRO Fellowships for Graduate Environmental Study (2004)
Research Category: Academic Fellowships , Ecological Indicators/Assessment/Restoration , Fellowship - Environmental Science/Forensic Science

Description:

Objective:

The objective of this research project is to investigate expression from the promoter regions of tfdR and tfdS coding for the regulatory proteins of the 2,4-dichlorophenoxyacetic acid (2,4-D) degradation pathway.

Approach:

The work can be divided into four steps. First, the promoter regions from the soil bacterium Ralstonia eutropha JMP134 will be cloned, and promoter lacZ fusions formed. Second, expression from both promoter regions in strain JMP134, the parent microorganism, will be tested under induced, uninduced, and stress conditions. Third, the promoter fusions will be transferred to a collection of 2,4-D degrading organisms and tested for levels of expression. Fourth, fusions will be compared and analyzed by qualitative and quantitative biochemical assays for differences in expression levels.

It is known that tfdR and tfdS both are able to regulate gene expression (Wright and Snyder, 1997); however, it may be that one is expressed to a greater level than the other or that one is expressed in a regulated fashion. This project should determine if one of the regulatory genes has a greater role in pathway regulation. The results also should provide clues as to the evolution of microbial regulatory systems of catabolic pathways, essential information for understanding the eventual fate of xenobiotic compounds in the environment. We expect expression from these promoters will not be highly regulated, though expression from the promoters of tfdR and tfdS may vary.

Supplemental Keywords:

fellowship, 2,4-dichlorophenoxyacetic acid, 2,4-D, promoter regions, tfdR, tfdS, Ralstonia eutropha JMP134, lacZ, gene regulation, xenobiotics, catabolic pathway, , Ecosystem Protection/Environmental Exposure & Risk, POLLUTANTS/TOXICS, Scientific Discipline, RFA, Ecosystem/Assessment/Indicators, exploratory research environmental biology, Ecological Indicators, Biochemistry, Environmental Microbiology, Ecological Effects - Environmental Exposure & Risk, Genetics, Microorganisms, bacteria, xenobiotics, microbial communities, 2, 4-Dichlorophenoxyacetic, degradation, genetic engineering, biochemical indicators, genetic differentiation, genetically engineered microorganisms

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The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Conclusions drawn by the principal investigators have not been reviewed by the Agency.

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