Detection and quantification of expression of amoA by competitive reverse transcription-PCR

Y. Ebie*, H. Miura, N. Noda, M. Matsumura, S. Tsuneda, A. Hirata, Y. Inamori

*この研究の対応する著者

研究成果: Article査読

5 被引用数 (Scopus)

抄録

Ammonia oxidation by chemolithoautotrophic ammonia-oxidizing bacteria is an important step in the biological nitrogen removal process. The first conversion step, the oxidation of ammonia to hydroxylamine is catalyzed by ammonia monooxygenase (AMO). To investigate the activity of ammonia oxidation, mRNA (designated as amoA) encoding a subunit of AMO was quantified by competitive reverse transcription (RT)-PCR. As a result, it was possible to detect and quantify amoA expression in cultured Nitrosomonas europaea and even complex microbial communities such as nitrifying bacterial aggregates by competitive RT-PCR. It was estimated that amoA concentration in cultured N. europaea was 2.3 × 108 copies.ml-1. Additionally, it was calculated that the copy number of amoA in nitrifying bacterial aggregates was 1.0 × 1012 copies.ml-1 (5.1 × 1010 copies.mg-1-dry weight). On the other hand, amoA expression in the natural activated sludge in a household Gappei-Johkaso was undetectable, whereas 16S rRNA of ammonia-oxidizing bacteria was detected by RT-PCR. Then, four days cultivation of this sludge in inorganic artificial wastewater resulted in increasing amoA expression to a quantifiable amount by competitive RT-PCR. In conclusion, the competitive RT-PCR was effective to investigate the expression of amoA as an indicator of ammonia oxidation activity by autotrophic ammonia-oxidizing bacteria.

本文言語English
ページ(範囲)281-288
ページ数8
ジャーナルWater Science and Technology
46
1-2
DOI
出版ステータスPublished - 2002
外部発表はい

ASJC Scopus subject areas

  • 環境工学
  • 水の科学と技術

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