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MB Sample ID: SA176604
| Local Sample ID: | OMV-1526-1 |
| Subject ID: | SU001982 |
| Subject Type: | Bacteria |
| Subject Species: | Bacteroides thetaiotaomicron |
| Taxonomy ID: | 818 |
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Collection:
| Collection ID: | CO001975 |
| Collection Summary: | B. thetaiotaomicron strains (wild-type and ΔBT1522-BT1526 mutants) were grown overnight in an anaerobic chamber (Coy Laboratories) using an atmosphere of 10% H2, 5% CO2, 85% N2. For liquid growth, Brain heart infusion (BHI) supplemented with hemin and vitamin K3 was used. Cultures were then subjected to subcellular fractionation to obtain total membranes and outer membrane vesicles (OMV) preparation. OMV preparations: OMV were purified by ultracentrifugation of filtered spent media from 150 ml of liquid culture as described previously (1). OMV preparations were resuspended in PBS before lipid analyses. Protein content was quantified using a DC protein assay kit (Bio-Rad). Fractions were aliquoted and stored at -80°C until analyzed. Membrane preparations: Total membrane preparations were performed by cell lysis and ultracentrifugation as previously described (1). Total membranes from 150 ml of liquid culture were resuspended in PBS using a 2-ml glass tissue grinder with a polytetrafluoroethylene (PTFE) pestle (VWR). Protein content was quantified using a DC protein assay kit (Bio-Rad). Fractions were stored aliquoted and stored at -80°C until analyzed. References: 1. Elhenawy W, Debelyy MO, Feldman MF. Preferential packing of acidic glycosidases and proteases into Bacteroides outer membrane vesicles. mBio. 2014;5(2):e00909-14. |
| Sample Type: | Bacterial cells |
