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dc.contributor.authorReichart, Nicholas J.
dc.contributor.authorJay, Zachary J.
dc.contributor.authorKrukenberg, Viola
dc.contributor.authorParker, Albert E.
dc.contributor.authorSpietz, Rachel L.
dc.contributor.authorHatzenpichler, Roland
dc.date.accessioned2022-04-14T19:45:46Z
dc.date.available2022-04-14T19:45:46Z
dc.date.issued2020
dc.identifier.citationNicholas J. Reichart, Zackary J. Jay, Viola Krukenberg, Albert E. Parker, Rachel L. Spietz, Roland Hatzenpichler (2020). Minimum information guideline for spectrophotometric and fluorometric methods to assess biofilm formation in microplates. The ISME Journal, 14(11), 2851-2861. 10.1038/s41396-020-00749-1en_US
dc.identifier.issn1751-7362
dc.identifier.urihttps://scholarworks.montana.edu/xmlui/handle/1/16726
dc.description.abstractMetagenomic studies have revolutionized our understanding of the metabolic potential of uncultured microorganisms in various ecosystems. However, many of these genomic predictions have yet to be experimentally tested, and the functional expression of genomic potential often remains unaddressed. In order to obtain a more thorough understanding of cell physiology, novel techniques capable of testing microbial metabolism under close to in situ conditions must be developed. Here, we provide a benchmark study to demonstrate that bioorthogonal non-canonical amino acid tagging (BONCAT) in combination with fluorescence-activated cell sorting (FACS) and 16S rRNA gene sequencing can be used to identify anabolically active members of a microbial community incubated in the presence of various growth substrates or under changing physicochemical conditions. We applied this approach to a hot spring sediment microbiome from Yellowstone National Park (Wyoming, USA) and identified several microbes that changed their activity levels in response to substrate addition, including uncultured members of the phyla Thaumarchaeota, Acidobacteria, and Fervidibacteria. Because shifts in activity in response to substrate amendment or headspace changes are indicative of microbial preferences for particular growth conditions, results from this and future BONCAT-FACS studies could inform the development of cultivation media to specifically enrich uncultured microbes. Most importantly, BONCAT-FACS is capable of providing information on the physiology of uncultured organisms at as close to in situ conditions as experimentally possible.en_US
dc.language.isoen_USen_US
dc.publisherSpringer Science and Business Media LLCen_US
dc.rightsThis article is made available under the CC-BY 4.0 licenseen_US
dc.titleActivity-based cell sorting reveals responses of uncultured archaea and bacteria to substrate amendmenten_US
dc.typeArticleen_US
mus.citation.extentfirstpage2851en_US
mus.citation.extentlastpage2861en_US
mus.citation.issue11en_US
mus.citation.journaltitleThe ISME Journalen_US
mus.citation.volume14en_US
mus.identifier.doi10.1038/s41396-020-00749-1en_US
mus.relation.collegeCollege of Engineeringen_US
mus.relation.collegeCollege of Letters & Scienceen_US
mus.relation.departmentCenter for Biofilm Engineering.en_US
mus.relation.departmentChemistry & Biochemistry.en_US
mus.relation.departmentMathematical Sciences.en_US
mus.relation.universityMontana State University - Bozemanen_US
mus.relation.researchgroupCenter for Biofilm Engineering.en_US
mus.relation.researchgroupThermal Biology Institute (TBI).en_US


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