An array of transporter knockout mutants of the marine bacterium Ruegeria pomeroyi DSS-3 mutants were screened for growth on 70 environmentally relevant marine substrates. The array contains 156 isolated knockout mutants of putative transporter genes as well as positive and negative growth controls and is distributed across two 96-well plates. Actively growing cultures were inoculated into freshly prepared 96 well plates containing minimal medium with a single substrates as sole carbon source. P...
Show moreThe data were generated from laboratory experiments performed at the University of Florida, Gainesville, Florida.
Mutant cultures were pre-grown overnight in ½ YTSS medium with 50 mg ml-1 kanamycin. Screens were performed in L1 minimal medium (Guillard and Hargraves 1993) modified to a salinity of 20 ppt and amended with ammonium (3 mM), kanamycin (50 mg ml-1), and phosphorus as PO43- at 36 mM. Overnight cultures of individual mutants (2 ml) were inoculated into 198 ml of modified L1 with a single substrate as the sole carbon source at 8 mM carbon. Plates were incubated at 25oC with shaking, and optical density (OD600) was read at intervals of 6-24 h until cultures entered stationary phase at ~24-72 h (SpectraMax M3, Molecular Devices, San Jose, CA). Mutants exhibiting phenotypes in the initial screen were moved to the targeted screen.
Schroer, W. F., Moran, M. A., Reisch, C. R., Mejia, C., Trujillo Rodriguez, L., Gray, L. G. (2023) High throughput growth screening of the marine bacterium Ruegeria pomeroyi DSS-3 knockout mutants on 70 environmentally relevant marine substrates done in 2021. Biological and Chemical Oceanography Data Management Office (BCO-DMO). (Version 1) Version Date 2023-07-17 [if applicable, indicate subset used]. doi:10.26008/1912/bco-dmo.904246.1 [access date]
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