Heterogeneous expression of Pil1 depends on its 5′ upstream region.


<p>(A–D) Flow cytometry profiles of the WT UCN34 strain, the isogenic Δ<i>pil1</i> mutant, and the Δ<i>pil1</i> complemented with pTCV<i>erm</i>-P<i>tet</i>-<i>pil1</i> or pTCV<i>erm</i>-P<i>pil1</i>-<i>pil1</i>. (A) The WT strain displays two cell subpopulations, a majority of Pil1<sub>low</sub> (66%) and a minority of Pil1<sub>high</sub> (27%). (B) The Δ<i>pil1</i> mutant that does not express the Pil1 pilus. (C) The Δ<i>pil1</i> mutant complemented with the plasmid pTCV<i>erm</i>-P<i>tet</i>-<i>pil1</i>, P<i>tet</i> being a constitutive promoter, displays a single population of highly piliated cells. (D) The Δ<i>pil1</i> mutant complemented with the plasmid pTCV<i>erm</i>-P<i>pil1</i>-<i>pil1</i>, P<i>pil1</i> being the entire <i>gallo2180-pilA</i> intergenic region, displays two subpopulations. (C–D) Immunofluorescence staining with anti-PilB pAb (green) confirms that only the P<i>pil1</i> promoter and downstream sequences restored the heterogeneous expression of the Pil1 pilus (Pil1<sub>low</sub>, 76%; Pil1<sub>high</sub>, 16%). Bacteria were stained with DAPI (blue).</p

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