Draft Genome Sequence of the Methane-Oxidizing Bacterium “
            <i>Candidatus</i>
            Methylomonas sp. LWB” Isolated from Movile Cave

Stephenson, Jason R.; Kumaresan, Deepak; Hillebrand-Voiculescu, Alexandra; Brooks, Elliot; Whiteley, Andrew S.; Murrell, J. Colin

doi:10.1128/genomea.01491-16

Cited by 4 publications

(1 citation statement)

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“…In the latter cases, more phylum specific primers had to be designed to investigate the presence of ‘ Candidatus Methylomirabilis oxyfera’ in various ecosystems (Luesken et al 2011b ). Recently several genomes of different MOB have been sequenced by the Omega consortium (Khmelenina et al 2013 ; Kits et al 2013 ; Khadem et al 2012 ; Stephenson et al 2017 ) and thus a much larger gene dataset is now available to design new primers to potentially cover a larger methanotroph diversity. Here we introduce a new set of degenerate primers that can be used to examine the diversity of MOB in various environments with the potential ability to target all presently known methanotrophic phyla.…”

Section: Introductionmentioning

confidence: 99%

Survey of methanotrophic diversity in various ecosystems by degenerate methane monooxygenase gene primers

2017

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Methane is the second most important greenhouse gas contributing to about 20% of global warming. Its mitigation is conducted by methane oxidizing bacteria that act as a biofilter using methane as their energy and carbon source. Since their first discovery in 1906, methanotrophs have been studied using a complementary array of methods. One of the most used molecular methods involves PCR amplification of the functional gene marker for the diagnostic of copper and iron containing particulate methane monooxygenase. To investigate the diversity of methanotrophs and to extend their possible molecular detection, we designed a new set of degenerate methane monooxygenase primers to target an 850 nucleotide long sequence stretch from pmoC to pmoA. The primers were based on all available full genomic pmoCAB operons. The newly designed primers were tested on various pure cultures, enrichment cultures and environmental samples using PCR. The results demonstrated that this primer set has the ability to correctly amplify the about 850 nucleotide long pmoCA product from Alphaproteobacteria, Gammaproteobacteria, Verrucomicrobia and the NC10 phyla methanotrophs. The new primer set will thus be a valuable tool to screen ecosystems and can be applied in conjunction with previously used pmoA primers to extend the diversity of currently known methane-oxidizing bacteria.Electronic supplementary materialThe online version of this article (doi:10.1186/s13568-017-0466-2) contains supplementary material, which is available to authorized users.

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