Detection of genes for periplasmic nitrate reductase in nitrate respiring bacteria and in community DNA

Flanagan, Deborah A., Gregory, Lisa G., Carter, Jon P., Karakas-Sen, Asuman, Richardson, David J. ORCID: https://orcid.org/0000-0002-6847-1832 and Spiro, Stephen (1999) Detection of genes for periplasmic nitrate reductase in nitrate respiring bacteria and in community DNA. FEMS Microbiology Letters, 177 (2). pp. 263-270. ISSN 0378-1097

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Abstract

A nested PCR primed by four degenerate oligonucleotides was developed for the specific amplification of sequences from the napA gene encoding the periplasmic nitrate reductase. This approach was used to amplify fragments of the napA gene from 10 Pseudomonas species and one Moraxella sp., previously shown to be able to express the periplasmic nitrate reductase activity, from Rhodobacter capsulatus and from community DNA extracted from a fresh-water sediment. Amino acid sequences encoded by the napA fragments were compared to one another and to the corresponding regions of related enzymes. This comparison indicates that the amplification protocol is specific for its intended target. The napA sequences amplified from community DNA were tightly clustered, which may indicate a degree of homogeneity in the sediment community. All tested Gram-negative strains capable of aerobic nitrate respiration were found to have periplasmic nitrate reductase genes. However, some strains which have and express the genes are incapable of aerobic nitrate respiration. The PCR primers and amplification protocols described will be useful in future studies of nitrate respiring populations.

Item Type: Article
Additional Information: Funding Information: This work was supported by BBSRC and Royal Society grants to DJR and SS, NERC studentships to DAF and LGG and a Turkish Government grant to AKS. We are grateful to Dr. R. James for synthesising oligonucleotides and to N. Shearer and Dr. B. Berks for helpful discussions.
Uncontrolled Keywords: denitrification,nitrate respiration,periplasmic nitrate reductase,microbiology,molecular biology,genetics ,/dk/atira/pure/subjectarea/asjc/2400/2404
Faculty \ School:

Faculty of Science > School of Biological Sciences
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Depositing User: LivePure Connector
Date Deposited: 15 Jul 2022 11:30
Last Modified: 12 Aug 2022 05:36
URI: https://ueaeprints.uea.ac.uk/id/eprint/86185
DOI: 10.1016/S0378-1097(99)00324-9

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