Growth of E. coli wild-type (wt) and entE strains in liquid and solid media (original) (raw)
2014
Abstract
<p>A) Liquid aerated minimal M9 medium cultures of wild-type strain (blue squares), <i>entE</i> strain (green circles) and <i>entE</i> strain in the same media but supplemented with 100 µM FeCl<sub>3</sub> (red triangles). Growth (OD<sub>600</sub>) was determined at the indicated times. B) Lawn growth of wt and <i>entE E. coli</i> strains on M9A. A stationary phase culture of <i>entE E. coli</i> strain was serially diluted (10<sup>−1</sup> to 10<sup>−4</sup>) and an aliquot of these dilutions was applied on M9A or M9A supplemented with 100 µM FeCl<sub>3</sub>. As control, the same dilutions of a wt strain overnight culture were applied on M9A medium. Lawn growth was compared at 8 hours of incubation. C) Colony growth of wt and <i>entE E. coli</i> on LBA. A stationary phase culture of <i>entE E. coli</i> strain was serially diluted and an aliquot of dilutions 10<sup>−6</sup> to 10<sup>−8</sup> were applied on LBA or LBA supplemented with 100 µM FeCl<sub>3</sub>. As control, the same dilutions of an overnight culture of the wt strain were applied on LBA medium. After overnight incubation, colonies sizes were compared. D) Activity of the <i>rhyB</i> promoter estimated by β-galactosidase activity as an indirect measure of the intracellular iron content (The higher the promoter expression, the lower the iron content <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0084734#pone.0084734-Ma2" target="_blank">[48]</a>). Both wild-type strain and <i>entE</i> mutant respond to iron addition. The plasmid pALM23 carries the <i>ryhB- lacZ</i> fusion.</p
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