Factors Influencing Production of the Antibiotic Pyoluteorin in Pseudomonas Fluorescens Pf-5EPA Grant Number: U915828
Title: Factors Influencing Production of the Antibiotic Pyoluteorin in Pseudomonas Fluorescens Pf-5
Investigators: Brodhagen, Marion L.
Institution: Oregon State University
EPA Project Officer: Manty, Dale
Project Period: June 1, 2000 through June 1, 2003
Project Amount: $101,703
RFA: STAR Graduate Fellowships (2000) RFA Text | Recipients Lists
Research Category: Fellowship - Molecular Biology/Genetics , Academic Fellowships , Biology/Life Sciences
The objective of this research is to characterize transcriptional regulation of the pyoluteorin (PLT) biosynthetic genes in Pseudomonas fluorescens Pf-5 by the pathway-linked regulatory genes pltR and pltH, and to determine whether global regulatory circuits that control PLT biosynthetic gene transcription involve PltR and PltH.
Northern analysis and primer extension will be used to define the promoters within the PLT biosynthetic gene cluster. DNA footprinting analysis will be used to test suspected protein-promoter interactions. Global regulatory genes have been observed to influence PLT biosynthetic gene transcription both positively (gacA and gacS, which together encode a two-component regulatory system) and negatively (lon, which encodes a serine protease, and rpoS, which encodes the stationary phase sigma factor). Comparisons between wild-type Pf-5 and strains harboring mutations in each of these global regulatory genes will be made, to detect differences between them in both transcriptional activity and accumulation of protein products from pltR and pltH. Northern and western analysis will be used to measure transcript and protein accumulation, respectively.
This study is expected to increase our understanding of how to improve performance of microbial biological control agents.