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Record 2 of 1139
Parallel characterization of anaerobic toluene- and ethylbenzene-degrading microbial consortia by PCR-denaturing gradient gel electrophoresis, RNA-DNA membrane hybridization, and DNA microarray technology
External Online Source: doi:10.1128/AEM.68.7.3215-3225.2002
Author and Affiliation:
Koizumi, Yoshikazu(Graduate School of Science, Tokyo Metropolitan University, Department of Biological Sciences, 1-1 Minami-Ohsawa, Hachioji, Tokyo 192-0397, Japan)
Kelly, John J.
Nakagawa, Tatsunori
Urakawa, Hidetoshi
El-Fantroussi, Said
Al-Muzaini, Saleh
Fukui, Manabu
Urushigawa, Yoshikuni
Stahl, David A.
Abstract: A mesophilic toluene-degrading consortium (TDC) and an ethylbenzene-degrading consortium (EDC) were established under sulfate-reducing conditions. These consortia were first characterized by denaturing gradient gel electrophoresis (DGGE) fingerprinting of PCR-amplified 16S rRNA gene fragments, followed by sequencing. The sequences of the major bands (T-1 and E-2) belonging to TDC and EDC, respectively, were affiliated with the family Desulfobacteriaceae. Another major band from EDC (E-1) was related to an uncultured non-sulfate-reducing soil bacterium. Oligonucleotide probes specific for the 16S rRNAs of target organisms corresponding to T-1, E-1, and E-2 were designed, and hybridization conditions were optimized for two analytical formats, membrane and DNA microarray hybridization. Both formats were used to characterize the TDC and EDC, and the results of both were consistent with DGGE analysis. In order to assess the utility of the microarray format for analysis of environmental samples, oil-contaminated sediments from the coast of Kuwait were analyzed. The DNA microarray successfully detected bacterial nucleic acids from these samples, but probes targeting specific groups of sulfate-reducing bacteria did not give positive signals. The results of this study demonstrate the limitations and the potential utility of DNA microarrays for microbial community analysis.
Publication Date: Jul 01, 2002
Document ID:
20040088245
(Acquired Sep 07, 2004)
Subject Category: LIFE SCIENCES (GENERAL)
Document Type: Journal Article
Publication Information: Applied and environmental microbiology (ISSN 0099-2240); Volume 68; 7; 3215-25
Publisher Information: United States
Description: In English
Distribution Limits: Unclassified; Publicly available; Unlimited
Rights: Copyright
NASA Terms: BACTERIA; DEGRADATION; DEOXYRIBONUCLEIC ACID; ELECTROPHORESIS; GELS; GRADIENTS; MEMBRANES; MICROORGANISMS; ORGANIZATIONS; RIBONUCLEIC ACIDS; TOLUENE; CLASSIFICATIONS; GENETICS; NUCLEIC ACIDS; OLIGONUCLEOTIDES; POLYMERASE CHAIN REACTION; SULFIDES
Other Descriptors: BACTERIA/CLASSIFICATION/GENETICS/METABOLISM; BENZENE DERIVATIVES/METABOLISM; TOLUENE/METABOLISM; MOLECULAR SEQUENCE DATA; NUCLEIC ACID HYBRIDIZATION; OLIGONUCLEOTIDE ARRAY SEQUENCE ANALYSIS; POLYMERASE CHAIN REACTION; SEQUENCE ANALYSIS, DNA; SULFIDES/METABOLISM; SUPPORT, NON-U.S. GOV'T; SUPPORT, U.S. GOV'T, NON-P.H.S; NASA DISCIPLINE LIFE SCIENCES TECHNOLOGIES; NON-NASA CENTER
Availability Source: Other Sources
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