| Title: | "Quantification of homogentisate-1,2-dioxygenase expression in a fungus degrading ethylbenzene" |
| Author(s): | Gunsch CK; Kinney KA; Szaniszlo PJ; Whitman CP; |
| Address: | "Department of Civil and Environmental Engineering, Duke University, United States" |
| DOI: | 10.1016/j.mimet.2006.03.018 |
| ISSN/ISBN: | 0167-7012 (Print) 0167-7012 (Linking) |
| Abstract: | "A quantitative real time reverse transcription polymerase chain reaction (qRT-PCR) assay was utilized to quantify the expression of ElHDO in the fungus Exophiala lecanii-corni during the biodegradation of ethylbenzene and other volatile organic pollutants. The assay was applied to measure the impact of pollutant mixtures on ElHDO expression relative to that of a housekeeping gene (18S rRNA). Three compounds were tested in mixtures with ethylbenzene: methyl propyl ketone, phenylacetate and o-xylene. These chemicals repressed, induced, or had no effect on ethylbenzene degradation, respectively. The results demonstrate that the gene target expression value (T(N)) is a useful parameter for evaluating the effect of pollutant mixtures on gene expression. T(N) was found to reflect macroscopic changes in ethylbenzene utilization rates although these two parameters were not related in a linear fashion for all compounds. The assay was log-linear over 5 orders of magnitude of RNA concentration and reproducible between samples (the largest T(N) standard deviation was 20%). The comparative qRT-PCR assay used in this research represents a viable alternative to absolute quantification methods to monitor in situ fungal gene expression in natural and engineered environmental systems" |
| Keywords: | "Benzene Derivatives/*metabolism Biodegradation, Environmental Exophiala/*enzymology/genetics Homogentisate 1, 2-Dioxygenase/blood/genetics/*metabolism Pentanones/metabolism Phenylacetates/metabolism Reverse Transcriptase Polymerase Chain Reaction Water Pol;" |
| Notes: | "MedlineGunsch, Claudia K Kinney, Kerry A Szaniszlo, Paul J Whitman, Christian P eng Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Netherlands 2006/05/17 J Microbiol Methods. 2006 Nov; 67(2):257-65. doi: 10.1016/j.mimet.2006.03.018. Epub 2006 May 15" |
