{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T19:17:28Z","timestamp":1790623048720,"version":"4.1.0"},"reference-count":34,"publisher":"American Society for Microbiology","issue":"19","license":[{"start":{"date-parts":[[2016,9,16]],"date-time":"2016-09-16T00:00:00Z","timestamp":1473984000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2016,10,1]],"date-time":"2016-10-01T00:00:00Z","timestamp":1475280000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Appl Environ Microbiol"],"published-print":{"date-parts":[[2016,10]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            Two genes,\n            <jats:italic>hgcA<\/jats:italic>\n            and\n            <jats:italic>hgcB<\/jats:italic>\n            , are essential for microbial mercury (Hg) methylation. Detection and estimation of their abundance, in conjunction with Hg concentration, bioavailability, and biogeochemistry, are critical in determining potential hot spots of methylmercury (MeHg) generation in at-risk environments. We developed broad-range degenerate PCR primers spanning known\n            <jats:italic>hgcAB<\/jats:italic>\n            genes to determine the presence of both genes in diverse environments. These primers were tested against an extensive set of pure cultures with published genomes, including 13\n            <jats:named-content content-type=\"genus-species\">Deltaproteobacteria<\/jats:named-content>\n            , nine\n            <jats:named-content content-type=\"genus-species\">Firmicutes<\/jats:named-content>\n            , and nine methanogenic\n            <jats:named-content content-type=\"genus-species\">Archaea<\/jats:named-content>\n            genomes. A distinct PCR product at the expected size was confirmed for all\n            <jats:italic>hgcAB<\/jats:italic>\n            <jats:sup>+<\/jats:sup>\n            strains tested via Sanger sequencing. Additionally, we developed clade-specific degenerate quantitative PCR (qPCR) primers that targeted\n            <jats:italic>hgcA<\/jats:italic>\n            for each of the three dominant Hg-methylating clades. The clade-specific qPCR primers amplified\n            <jats:italic>hgcA<\/jats:italic>\n            from 64%, 88%, and 86% of tested pure cultures of\n            <jats:named-content content-type=\"genus-species\">Deltaproteobacteria<\/jats:named-content>\n            ,\n            <jats:named-content content-type=\"genus-species\">Firmicutes<\/jats:named-content>\n            , and\n            <jats:named-content content-type=\"genus-species\">Archaea<\/jats:named-content>\n            , respectively, and were highly specific for each clade. Amplification efficiencies and detection limits were quantified for each organism. Primer sensitivity varied among species based on sequence conservation. Finally, to begin to evaluate the utility of our primer sets in nature, we tested\n            <jats:italic>hgcA<\/jats:italic>\n            and\n            <jats:italic>hgcAB<\/jats:italic>\n            recovery from pure cultures spiked into sand and soil. These novel quantitative molecular tools designed in this study will allow for more accurate identification and quantification of the individual Hg-methylating groups of microorganisms in the environment. The resulting data will be essential in developing accurate and robust predictive models of Hg methylation potential, ideally integrating the geochemistry of Hg methylation to the microbiology and genetics of\n            <jats:italic>hgcAB<\/jats:italic>\n            .\n          <\/jats:p>\n          <jats:p>\n            <jats:bold>IMPORTANCE<\/jats:bold>\n            The neurotoxin methylmercury (MeHg) poses a serious risk to human health. MeHg production in nature is associated with anaerobic microorganisms. The recent discovery of the Hg-methylating gene pair,\n            <jats:italic>hgcA<\/jats:italic>\n            and\n            <jats:italic>hgcB<\/jats:italic>\n            , has allowed us to design and optimize molecular probes against these genes within the genomic DNA for microorganisms known to methylate Hg. The protocols designed in this study allow for both qualitative and quantitative assessments of pure-culture or environmental samples. With these protocols in hand, we can begin to study the distribution of Hg-methylating organisms in nature via a cultivation-independent strategy.\n          <\/jats:p>","DOI":"10.1128\/aem.01271-16","type":"journal-article","created":{"date-parts":[[2016,7,16]],"date-time":"2016-07-16T03:19:05Z","timestamp":1468639145000},"page":"6068-6078","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":88,"title":["Development and Validation of Broad-Range Qualitative and Clade-Specific Quantitative Molecular Probes for Assessing Mercury Methylation in the Environment"],"prefix":"10.1128","volume":"82","author":[{"given":"Geoff A.","family":"Christensen","sequence":"first","affiliation":[{"name":"Biosciences Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ann M.","family":"Wymore","sequence":"additional","affiliation":[{"name":"Biosciences 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