Affiliation:
1. Department of Chemical Engineering and Applied Chemistry, University of Toronto
2. Department of Chemistry, Biology, and Chemical Engineering, Ryerson University, Toronto
3. Division of Physical and Environmental Sciences, University of Toronto, Scarborough, Ontario, Canada
Abstract
ABSTRACT
A DNA microarray to monitor the expression of bacterial metabolic genes within mixed microbial communities was designed and tested. Total RNA was extracted from pure and mixed cultures containing the 2,4-dichlorophenoxyacetic acid (2,4-D)-degrading bacterium
Ralstonia eutropha
JMP134, and the inducing agent 2,4-D. Induction of the 2,4-D catabolic genes present in this organism was readily detected 4, 7, and 24 h after the addition of 2,4-D. This strain was diluted into a constructed mixed microbial community derived from a laboratory scale sequencing batch reactor. Induction of two of five 2,4-D catabolic genes (
tfdA
and
tfdC
) from populations of JMP134 as low as 10
5
cells/ml was clearly detected against a background of 10
8
cells/ml. Induction of two others (
tfdB
and
tfdE
) was detected from populations of 10
6
cells/ml in the same background; however, the last gene,
tfdF
, showed no significant induction due to high variability. In another experiment, the induction of resin acid degradative genes was statistically detectable in sludge-fed pulp mill effluent exposed to dehydroabietic acid in batch experiments. We conclude that microarrays will be useful tools for the detection of bacterial gene expression in wastewaters and other complex systems.
Publisher
American Society for Microbiology
Subject
Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology
Reference35 articles.
1. Ausubel F. M. R. Brent R. E. Kingston D. D. Moore J. G. Seidman J. A. Smith and K. Struhl. 1987. Current protocols in molecular biology. John Wiley & Sons Inc. New York N.Y.
2. Bartlett, M. S. 1947. The use of transformations. Biometrics3:39-52.
3. Quantitative Detection of Microbial Genes by Using DNA Microarrays
4. de Saizieu, A., U. Certa, J. Warrington, C. Gray, W. Keck, and J. Mous. 1998. Bacterial transcript imaging by hybridization of total RNA to oligonucleotide arrays. Nat. Biotechnol.16:45-48.
5. Transposon mutagenesis and cloning analysis of the pathways for degradation of 2,4-dichlorophenoxyacetic acid and 3-chlorobenzoate in Alcaligenes eutrophus JMP134(pJP4)
Cited by
120 articles.
订阅此论文施引文献
订阅此论文施引文献，注册后可以免费订阅5篇论文的施引文献，订阅后可以查看论文全部施引文献