Nitrous oxide reductase (nosZ) gene-specific PCR primers for detection of denitrifiers and threenosZgenes from marine sediments
Open Access
- 1 May 1998
- journal article
- Published by Oxford University Press (OUP) in FEMS Microbiology Letters
- Vol. 162 (1), 61-68
- https://doi.org/10.1111/j.1574-6968.1998.tb12979.x
Abstract
Two PCR primer sets for the nitrous oxide reductase gene (nosZ) were developed. The initial primers were based on three sequences in GenBank and used to amplify nosZ from continental shelf sediments and from two denitrifiers in culture, Thiosphaera pantotropha and Pseudomonas denitrificans. Three unique marine sediment nosZ genes were identified and sequenced. The marine nosZ genes were most closely related to the nosZ genes of Paracoccus denitrificans or to Rhizobium meliloti. Alignment of all nosZ sequences currently available (n= 10) facilitated redesign of the PCR primers. Three new primer sets which amplify 1100 bp, 900 bp and 250 bp regions of the nosZ gene were designed and tested. The new primers robustly amplified nosZ fragments from samples in which the initial nosZ primers were only marginally successful.Keywords
This publication has 14 references indexed in Scilit:
- Estimating denitrification in North Atlantic continental shelf sedimentsBiogeochemistry, 1996
- Characterisation and amino acid sequence of cytochrome c‐550 from Thiosphaera pantotrophaEuropean Journal of Biochemistry, 1994
- Sequence and expression of the gene encoding the respiratory nitrous‐oxide reductase from Paracoccus denitrificansEuropean Journal of Biochemistry, 1993
- Transfer of Thiosphaera pantotropha to Paracoccus denitrificansInternational Journal of Systematic and Evolutionary Microbiology, 1993
- Derived amino acid sequences of the nosZ gene (respiratory N2O reductase) from Alcaligenes eutrophus, Pseudomonas aeruginosa and Pseudomonas stutzeri reveal potential copper‐binding residuesEuropean Journal of Biochemistry, 1992
- Basic local alignment search toolJournal of Molecular Biology, 1990
- CLUSTAL: a package for performing multiple sequence alignment on a microcomputerGene, 1988
- A small-scale five-hour procedure for isolating multiple samples of CsCl-purified DNA: Application to isolations from mammalian, insect, higher plant, algal, yeast, and bacterial sourcesAnalytical Biochemistry, 1986
- The role of fixed nitrogen in atmospheric photochemistryPhilosophical Transactions of the Royal Society of London. B, Biological Sciences, 1982
- Greenhouse Effects due to Man-Made Perturbations of Trace GasesScience, 1976