Transcriptomics

Dataset Information

6

Characterization of mycobacterial oligopeptide permease system


ABSTRACT: M. tuberculosis strain 1254 wild type (WT) and its isogenic delta Rv3662c-Rv3665c KO (opp KO) and complemented opp::opp were cultured in vitro, in liquid 7H9 media supplemented with OADC and 0.05% Tween 80, 0.5% glycerol, in a 150 mL volume, contained in roller bottles, incubated at 37oC. Growth was monitored as OD600 nm and recorded on a daily basis. In all experiments, the starting OD600 nm was 0.04-0.05. Aliquots were removed at each OD600 nm to make serial dilutions, plate onto 7H10 OADC agar plates for CFU counts. When cells reached OD600 nm of 0.3, 1.0, and 1.5, a 10-30 mL aliquot was taken, centrifuged 3 min at 3000 g, at room temperature, and the supernatant was discarded, with cell pellets being immediately frozen by inclusion into dry ice. Frozen cell pellets were stored at -80oC for later RNA isolation. In all experiments, RNA from WT was labeled with Cy5, and RNA from the opp KO was labeled with Cy3. In all experiments, the microarray comparsion used RNA from cells of the same growth stage, i.e., WT OD 0.3 vs opp KO OD 0.3, WT OD 1.0 vs opp KO OD 1.0, and WT OD 1.5 vs opp KO OD 1.5. When comparing the Complemented opp::opp vs the mutant opp KO, RNA from the later strain was Cy3-labeled. A cell type comparison design experiment design type compares cells of different type for example different cell lines. cell_type_comparison_design

ORGANISM(S): Mycobacterium tuberculosis  

SUBMITTER: Stanford Microarray Database   Mario A Flores-Valdez 

PROVIDER: E-GEOD-18426 | ArrayExpress | 2009-10-06

SECONDARY ACCESSION(S): GSE18426PRJNA118223

REPOSITORIES: GEO, ArrayExpress

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Publications

Mycobacterium tuberculosis modulates its cell surface via an oligopeptide permease (Opp) transport system.

Flores-Valdez Mario Alberto MA   Morris Rowan P RP   Laval Françoise F   Daffé Mamadou M   Schoolnik Gary K GK  

FASEB journal : official publication of the Federation of American Societies for Experimental Biology 20090811 12


Bacterial species utilize a vast repertoire of surface structures to interact with their surroundings and employ a number of strategies to reconfigure the cellular envelope according to specific stimuli. Gram-positive bacteria, exemplified by Streptomyces and Bacillus species, control production of some exposed molecules by importing oligopeptide signals via permeases (Opp). Such oligopeptides modulate intracellular signaling pathways. In this work, we functionally characterized an Opp of the hu  ...[more]

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