Detection and diversity of expressed denitrification genes in estuarine sediments after reverse transcription-PCR amplification from mRNA

Nogales, Balbina and Timmis, Kenneth N. and Nedwell, David B. and Osborn, A. M. (2002) Detection and diversity of expressed denitrification genes in estuarine sediments after reverse transcription-PCR amplification from mRNA. Applied and Environmental Microbiology, 68 (10). pp. 5017-5025. ISSN 0099-2240

Full content URL: http://www.scopus.com/inward/record.url?eid=2-s2.0...

Full text not available from this repository.

Item Type:Article
Item Status:Live Archive

Abstract

The expression of five denitrification genes coding for two nitrate reductases (narG and napA), two nitrite reductases (nirS and nirK), and nitrous oxide reductase (nosZ) was analyzed by reverse transcription (RT)-PCR of mRNA extracted from two sediment samples obtained in the River Colne estuary (United Kingdom), which receives high nitrogen inputs and for which high denitrification rates have been observed. The presence of all five genes in both sediment samples was confirmed by PCR amplification from extracted DNA prior to analysis of gene expression. Only nirS and nosZ mRNAs were detected; nirS was detected directly as an RT-PCR amplification product, and nosZ was detected following Southern blot hybridization. This indicated that active expression of at least the nirS and nosZ genes was occurring in the sediments at the time of sampling. Amplified nirS RT-PCR products were cloned and analyzed by sequencing, and they were compared with amplified nirS gene sequences from isolates obtained from the same sediments. A high diversity of nirS sequences was observed. Most of the cloned nirS sequences retrieved were specific to one site or the other, which underlines differences in the compositions of the bacterial communities involved in denitrifrification in the two sediments analyzed.

Keywords:Cloning, Denitrification, DNA, Enzymes, Estuaries, Nitrogen compounds, RNA, Sediments, Gene sequences, Genes, DNA, messenger RNA, oligonucleotide, RNA 16S, denitrification, estuarine sediment, gene expression, polymerase chain reaction, article, Escherichia coli, estuary, molecular cloning, nonhuman, nucleotide sequence, Ochrobactrum anthropi, Paracoccus, reverse transcription polymerase chain reaction, sediment, Bacteria, Blotting, Southern, Geologic Sediments, Nitrite Reductases, Nitrogen Fixation, Phylogeny, Polymerase Chain Reaction, Seawater, Variation (Genetics), United Kingdom, Bacteria (microorganisms), Ochrobactrum
Subjects:C Biological Sciences > C180 Ecology
C Biological Sciences > C500 Microbiology
Divisions:College of Science > School of Life Sciences
ID Code:8964
Deposited On:22 Apr 2013 11:59

Repository Staff Only: item control page