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dc.contributor.authorDelaney, Nigel Francis
dc.contributor.authorKaczmarek, Maria E.
dc.contributor.authorWard, Lewis M.
dc.contributor.authorSwanson, Paige Kathleen
dc.contributor.authorLee, Ming-Chun
dc.contributor.authorMarx, Christopher J
dc.date.accessioned2014-02-25T12:48:20Z
dc.date.issued2013
dc.identifier.citationDelaney, Nigel F., Maria E. Kaczmarek, Lewis M. Ward, Paige K. Swanson, Ming-Chun Lee, and Christopher J. Marx. 2013. Development of an optimized medium, strain and high-throughput culturing methods for methylobacterium extorquens. PLoS ONE 8(4): e62957.en_US
dc.identifier.issn1932-6203en_US
dc.identifier.urihttp://nrs.harvard.edu/urn-3:HUL.InstRepos:11743702
dc.description.abstractMethylobacterium extorquens strains are the best-studied methylotrophic model system, and their metabolism of single carbon compounds has been studied for over 50 years. Here we develop a new system for high-throughput batch culture of M. extorquens in microtiter plates by jointly optimizing the properties of the organism, the growth media and the culturing system. After removing cellulose synthase genes in M. extorquens strains AM1 and PA1 to prevent biofilm formation, we found that currently available lab automation equipment, integrated and managed by open source software, makes possible reliable estimates of the exponential growth rate. Using this system, we developed an optimized growth medium for M. extorquens using response surface methodologies. We found that media that used EDTA as a metal chelator inhibited growth and led to inconsistent culture conditions. In contrast, the new medium we developed with a PIPES buffer and metals chelated by citrate allowed for fast and more consistent growth rates. This new Methylobacterium PIPES (‘MP’) medium was also robust to large deviations in its component ingredients which avoided batch effects from experiments that used media prepared at different times. MP medium allows for faster and more consistent growth than other media used for M. extorquens.en_US
dc.description.sponsorshipOrganismic and Evolutionary Biologyen_US
dc.language.isoen_USen_US
dc.publisherPublic Library of Scienceen_US
dc.relation.isversionofdoi:10.1371/journal.pone.0062957en_US
dc.relation.hasversionhttp://www.ncbi.nlm.nih.gov/pmc/articles/PMC3639900/pdf/en_US
dash.licenseLAA
dc.subjectBiologyen_US
dc.subjectBiotechnologyen_US
dc.subjectGenetic Engineeringen_US
dc.subjectGenetically Modified Organismsen_US
dc.subjectGeneticsen_US
dc.subjectHeredityen_US
dc.subjectPhenotypesen_US
dc.subjectMicrobiologyen_US
dc.subjectBacteriologyen_US
dc.subjectBacterial Evolutionen_US
dc.subjectBacterial Physiologyen_US
dc.subjectMicrobial Evolutionen_US
dc.subjectMicrobial Growth and Developmenten_US
dc.subjectMicrobial Physiologyen_US
dc.titleDevelopment of an Optimized Medium, Strain and High-Throughput Culturing Methods for Methylobacterium extorquensen_US
dc.typeJournal Articleen_US
dc.description.versionVersion of Recorden_US
dc.relation.journalPLoS ONEen_US
dash.depositing.authorMarx, Christopher J
dc.date.available2014-02-25T12:48:20Z
dc.identifier.doi10.1371/journal.pone.0062957*
dash.contributor.affiliatedSwanson, Paige Kathleen
dash.contributor.affiliatedDelaney, Nigel Francis
dash.contributor.affiliatedLee, Ming-Chun
dash.contributor.affiliatedMarx, Christopher J


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