Phenotypic comparison of Δ mutation or overexpression of diguanylate cyclase on pellicle production and mRNA levels

<p><b>Copyright information:</b></p><p>Taken from "A cyclic-di-GMP receptor required for bacterial exopolysaccharide production"</p><p></p><p>Molecular Microbiology 2007;65(6):1474-1484.</p><p>Published online Jan 2007</p><p>PMCID:PMC2170427.</p><p>© 2007 The Authors; Journal compilation © 2007 Blackwell Publishing Ltd</p> A and B. Pellicle formation in static culture as revealed by crystal violet staining for (A) PA14, PA14Δ, PA14Δ and PA14Δ harbouring vector control or induced for expression of diguanylate cyclases , or ; (B) PA14Δ, PA14ΔΔ, PA14ΔΔ and PA14ΔΔ. C. Expression of was determined by measuring β-galactosidase levels using a reporter construct in strains PA14, PA14Δ, PA14Δ and PA14Δ harbouring vector control or induced for expression of diguanylate cyclases , or . D. Quantification of PEL polysaccharide by binding to Congo red was normalized to total protein for strains PA14 pMMB-, PA14Δ pMMB-, PA14Δ and PA14ΔΔ

Residues required for c-di-GMP binding are required for PEL polysaccharide synthesis

<p><b>Copyright information:</b></p><p>Taken from "A cyclic-di-GMP receptor required for bacterial exopolysaccharide production"</p><p></p><p>Molecular Microbiology 2007;65(6):1474-1484.</p><p>Published online Jan 2007</p><p>PMCID:PMC2170427.</p><p>© 2007 The Authors; Journal compilation © 2007 Blackwell Publishing Ltd</p> Complementation of the Δ mutation in PA14Δ pDN19- with pMMB containing full-length and indicated point mutations. Quantification of PEL polysaccharide binding to Congo red was normalized to total protein for each strain. Pellicle formation for each stain was tested in static culture and revealed by crystal violet staining. PelD and mutant derivatives were fused at their C-termini to HA tags. Total-cell extracts were prepared, loaded equally and separated on SDS-PAGE, transferred to PVDF and PelD-HA was detected with anti-HA antibody and chemiluminescence