Deletion of <i>sX13</i> derogates virulence gene expression.

Abstract

<p>(A) <i>Xcv</i> strains 85-10 (wt) and the <i>sX13</i> deletion mutant (Δ<i>sX13</i>) carrying the empty vector (pB) or the sX13 expression construct (p<i>sX13</i>) and strains additionally expressing HrpG* (p<i>hrpG*</i>) were incubated for 3.5 hours in <i>hrp</i>-gene inducing medium XVM2. Total protein extracts were analyzed by immunoblotting using antibodies directed against HrpF, HrcN, HrcJ and GroEL. The experiment was repeated twice with similar results. (B) <i>Xcv</i> 85-10 (wt), Δ<i>sX13</i> and Δ<i>sX13</i>+<i>sX13</i>_ch and strains additionally expressing HrpG* were incubated for 3.5 hours in <i>hrp</i>-gene inducing medium XVM2. Total protein extracts were analyzed by immunoblotting using antibodies directed against HrcJ and GroEL. The experiment was repeated twice with similar results. (C) Indicated genes were analyzed by qRT-PCR using RNA from cultures described in (B). The amount of each RNA in <i>Xcv</i> 85-10 was set to 1. Data points and error bars represent mean values and standard deviations obtained with three independent biological samples. Asterisks indicate statistically significant differences compared to wt (<i>t</i>-test; <i>P</i><0.03).</p