Restriction pattern analysis by high-performance liquid chromatography of PCR-amplified 16S rDNA from scum-forming bacteria in activated sludge
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- HIRAISHI AKIRA
- Department of Ecological Engineering, Toyohashi University of Technology Laboratory of Environmental Biotechnology, Konishi Co.
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- SHIN YONG KOOK
- Laboratory of Environmental Biotechnology, Konishi Co. Present address: Korean Collection for Type Cultures, Genetic Resources Center, Korea Research Institute of Bioscience & Biotechnology, Korea Institute of Science & Technology
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- TAKEUCHI JUN-ICHI
- Kosuge Sewage Treatment Plant, Sewege Bureau of Tokyo
書誌事項
- タイトル別名
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- Restriction Pattern Analysis by High-performance Liquid Chromatography of PCR-amplified 16S rDNA Fragments from Scum-forming Bacteria in Activated Sludge.
- Restriction Pattern Analysis by High-pe
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説明
High-performance liquid chromatography (HPLC) with an anion-exchange nonporous column was used to analyze restriction fragment length polymorphism (RFLP) of the 16S ribosomal DNA (rDNA) of scum-forming actinomycetes found in three different activated sludge plants. 16S rDNA fragments from washed scum samples were amplified by PCR and cloned into plasmid vectors. The 16S rDNA inserts in the plasmids were reamplified in vitro, cut with restriction endonucleases, and separated directly by HPLC. Compared to conventional agarose electrophoresis, the HPLC system used allowed better separation of fragments up to 500bp. The HPLC-RFLP patterns of the scum clones were compared with those of PCR-amplified 16S rDNAs from authentic strains and with computer-predicted restriction patterns of the available 16S rDNA data on the actinomycetes. 16S rDNA-RFLP comparisons indicated that the predominat scum-forming bacteria were Gordona amarae in two activated sludge plants and Rhodococcus sp. in the remaining one. These findings agreed well with results of lipoquinone profiling of the scum performed concurrently. The results of this study indicated that a combination of 16S rDNA-RFLP analysis and quinone profiling should provide a more powerful non-cultivating tool for the identification of scum-forming bacteria in activated sludge.
収録刊行物
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- Ｍｉｃｒｏｂｅｓ ａｎｄ ｅｎｖｉｒｏｎｍｅｎｔｓ
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Ｍｉｃｒｏｂｅｓ ａｎｄ ｅｎｖｉｒｏｎｍｅｎｔｓ 12 (3), 57-68, 1997
日本微生物生態学会 / 日本土壌微生物学会 / Taiwan Society of Microbial Ecology / 植物微生物研究会 / 極限環境微生物学会
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詳細情報 詳細情報について
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- CRID
- 1390001204346611584
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- NII論文ID
- 110001272800
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- NII書誌ID
- AA11173196
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- ISSN
- 13474405
- 13426311
- http://id.crossref.org/issn/13426311
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- NDL書誌ID
- 4315337
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- 本文言語コード
- en
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