Summary of Study ST002381
This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR001531. The data can be accessed directly via it's Project DOI: 10.21228/M8ST4T This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.
| Study ID | ST002381 |
| Study Title | Ruegeria pomeroyi transporter mutant substrate drawdown |
| Study Summary | The goal of this project was to identify bacterial transporters responsible for uptake of environmentally relevant marine metabolites. We used the model marine heterotrophic bacterium Ruegeria pomeroyi DSS-3, for which an arrayed library of single gene knockout mutants has been generated by selecting isolated from a barcoded transposon mutant library (BasSeq). Knockout mutants of putative transporters were grown on minimal medium with a single substrate as sole carbon source. Mutant defect was assessed by comparing the substrate drawdown of isolated mutants to drawdown by a pooled mutant library (BarSeq), a proxy for wildtype fitness. |
| Institute | University of Georgia |
| Laboratory | Moran Lab, Edison Lab |
| Last Name | Uchimiya |
| First Name | Mario |
| Address | 315 Riverbend Rd, Athens, GA, 30602, USA |
| mario.uchimiya@uga.edu | |
| Phone | (706) 542-8387 |
| Submit Date | 2022-11-16 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | fid |
| Analysis Type Detail | NMR |
| Release Date | 2022-12-27 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Treatment:
| Treatment ID: | TR002482 |
| Treatment Summary: | Mutants of Ruegeria pomeroyi DSS-3 were grown on minimal medium containing a single substrate as sole carbon source, they were screened for their ability to drawdown this target substrate. Screens were performed in L1 minimal medium modified to a salinity of 20 and amended with ammonium (3 mM) and kanamycin (50 ug ml-1). For each mutant-substrate pair, 3 replicate 220 µl cultures were prepared in 96 well plates by inoculating 3 ul of washed (3x) overnight mutant cultures into minimal medium containing the candidate substrate at 8 mM carbon concentration. Cultures were grown shaking at 25oC for 24 h or 36 h, depending on the growth rate supported by the carbon source. As a positive control, four wells with the same medium were inoculated with washed overnight cultures of the pooled-BarSeq library, used as a proxy for wild-type R. pomeroyi fitness but harboring a transposon/kanamycin resistance gene insertion. |
| Treatment Protocol Filename: | 3_Treatment protocol_UGA_mutant_Nov2022.docx |
