Terminal Restriction Pattern Analysis of 16S rRNA Genes for the Characterization of Bacterial Communities of Activated Sludge
スポンサーリンク
概要
- 論文の詳細を見る
A culture-independent molecular technique using terminal restriction fragment length polymorphism (T-RFLP) of 16S rRNA genes was applied to the characterization of bacterial communities of activated sludge taken from different municipal sewage treatment plants. 16S rDNA fragments from the bulk DNA of sludge were amplified by PCR with a Cy5-labeled forward primer corresponding to nucleotide positions 8 to 27 and a reverse primer complementary to positions 907 to 926 in the Escherichia coli numbering system. The 16S rDNAs thus obtained were digested with tetrameric restriction enzymes and analyzed using a Pharmacia automated Dna sequencer. A preliminary study on a model DNA mixture prepared from different bacterial species showed that the fluorescence intensity of terminal fragments (T-RFs) of 16S rDNAs amplified and detected was directly proportional to the 16S rRNA gene copy number rather than the amount of genomic DNA of each species present. 16S rDNA fragments amplified from the sludges and digested with HhaI usually generated at least 60 T-RFs, among which T-RFs of around 208 bp were the most abundant regardless of the time or area sampled. Southern blot hybridization with oligonucleotide probes specific to the 5' terminal regions of the 16S rDNA of different phylogenetic groups indicated that the T-RFs of around 208 bp were derived from members of the β subclass of the class Proteobacteria. Hybridization with a probe specific to the class Actinobacteria failed to detect any appreciable signal. These results did not agree fully with those obtained by quinone profiling. The usefulness and limitations of the T-RFLP method for monitoring bacterial population dynamics in activated sludge were discussed.
- 公益社団法人日本生物工学会の論文
- 2000-08-25
著者
-
HIRAISHI AKIRA
Department of Ecological Engineering, Toyohashi University of Technology
-
IWASAKI MITSURU
Department of Ecological Engineering, Toyohashi University of Technology
-
Hiraishi Akira
Department Of Biology Faculty Of Science Tokyo Metropolitan University
-
SHINJO HISASHI
Ebara Research Institute
-
Iwasaki Mitsuru
Department Of Ecological Engineering Toyohashi University Of Technology
関連論文
- Phylogenetic and Transcriptional Analyses of a Tetrachloroethene-Dechlorinating "Dehalococcoides" Enrichment Culture TUT2264 and Its Reductive-Dehalogenase Genes
- Combined Use of Cyanoditolyl Tetrazolium Staining and Flow Cytometry for Detection of Metabolically Active Bacteria in a Fed-batch Composting Process
- Removal of polychlorinated dioxins by semi-aerobic fed-batch composting with biostimulation of "Dehalococcoides"(ENVIRONMENTAL BIOTECHNOLOGY)
- A Modified Cyanoditolyl Tetrazolium Reduction Method for Differential Detection of Metabolically Active Gram-positive and Gram-negative Bacteria
- An Improved Redox Dye-Staining Method Using 5-Cyano-2,3-Ditoryl Tetrazolium Chloride for Detection of Metabolically Active Bacteria in Activated Sludge
- Effects of Chemical Uncouplers on Microbial Biomass Production, Metabolic Activity, and Community Structure in an Activated Sludge System
- Activity and Community Composition of Denitrifying Bacteria in Poly(3-hydroxybutyrate-co-3-hydroxyvalerate)-Using Solid-phase Denitrification Processes
- Characterization of the Microbial Community and Culturable Denitrifying Bacteria in a Solid-phase Denitrification Process Using Poly(ε-caprolactone) as the Carbon and Energy Source
- B-21 Comamonadacae : the important poly-β-hydroxybutyrate-degrading denitrifiers in activated sludge
- Rare Bacterium of New Genus Isolated with Prolonged Enrichment Culture
- Enrichment and Phylogenetic Analysis of Moderately Thermophilic Myxobacteria from Hot Springs in Japan
- Significance of Lipoquinones as Quantitative Biomarkers of Bacterial Populations in the Environment
- High Culturability of Bacteria in Commercially Available Personal Composters for Fed-batch Treatment of Household Biowaste
- Microbiology of Fed-batch Composting
- Nucleotide Sequences of Genes Coding for Photosynthetic Reaction Centers and Light-Harvesting Proteins of Acidiphilium rubrum and Related Aerobic Acidophilic Bacteria
- Biodiversity of Dehalorespiring Bacteria with Special Emphasis on Polychlorinated Biphenyl/Dioxin Dechlorinators
- In Situ Detection and Identification of Microorganisms at Single-Colony Resolution by Spectral Imaging
- Water Availability Is a Critical Determinant of a Population Shift from Proteobacteria to Actinobacteria during Start-Up Operation of Mesophilic Fed-Batch Composting
- Activity and Phylogenetic Composition of Proteolytic Bacteria in Mesophilic Fed-batch Garbage Composters
- Isolation and Characterization of Phototrophic Purple Nonsulfur Bacteria from Chloroflexus and Cyanobacterial Mats in Hot Springs
- Biotransformation of Polychlorinated Dioxins and Microbial Community Dynamics in Sediment Microcosms at Different Contamination Levels
- Estimation of "Dehalococcoides" Populations in Lake Sediment Contaminated with Low Levels of Polychlorinated Dioxins
- Combined Use of Cyanoditolyl Tetrazolium Staining and Flow Cytometry for Detection of Metabolically Active Bacteria in a Fed-batch Composting Process
- Distribution of Dibenzofuran-Degrading Bacteria in Soils Polluted with Different Levels of Polychlorinated Dioxins
- A Great Leap forward in Microbial Ecology
- Population Dynamics and Acetate Utilization Kinetics of Two Different Species of Phototrophic Purple Nonsulfur Bacteria in a Continuous Co-culture System
- Distribution and Capacity for Utilization of Lower Fatty Acids of Phototrophic Purple Nonsulfur Bacteria in Wastewater Environments
- Effects of Organic Nutrient Strength on the Purple Nonsulfur Bacterial Content and Metabolic Activity of Photosynthetic Sludge for Wastewater Treatment
- Restriction Pattern Analysis by High-performance Liquid Chromatography of PCR-amplified 16s rDNA Fragments from Scum-forming Bacteria in Activated Sludge
- Differences in Phototrophic Growth on High Phosphate Concentrations among Rhodopseudomonas species
- A New Approach to Numerical Analyses of Microbial Quinone Profiles in the Environment
- Terminal Restriction Pattern Analysis of 16S rRNA Genes for the Characterization of Bacterial Communities of Activated Sludge
- Isoprenoid Quinones as Biomarkers of Microbial Populations in the Environment
- A Great Leap forward in Microbial Ecology
- Isolation and Functional Gene Analyses of Aromatic-Hydrocarbon-Degrading Bacteria from a Polychlorinated-Dioxin-Dechlorinating Process
- Nitrate Removal Efficiency and Bacterial Community Dynamics in Denitrification Processes Using Poly (L-lactic acid) as the Solid Substrate
- Biodiversity of Dioxin-Degrading Microorganisms and Potential Utilization in Bioremediation
- Anaerobic Degradation of Aromatic Compounds by Magnetospirillum Strains: Isolation and Degradation Genes
- Isolation and Functional Gene Analyses of Aromatic-Hydrocarbon-Degrading Bacteria from a Polychlorinated-Dioxin-Dechlorinating Process