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. 2013 May 15;27(10):1132–1145. doi: 10.1101/gad.214734.113

Figure 6.

Figure 6.

PGA synthesis and biofilm formation with McaS mutants. (A) The MG1655 ΔmcaS ΔrpoS strain carrying either pNDM-McaS, pNDM-McaS-8, or pNDM-McaS-3 and its ΔpgaA derivative harboring pNDM-McaS were grown on Congo red indicator plates with 5 μM IPTG to assay PGA production. (B) NM525 ΔabgR-ydaL carrying pBR, pBR-McaS, pBR-McaS-3, or pBR-McaS-8 and NM525 ΔabgR-ydaL ΔpgaA∷cat carrying pBR-McaS were grown for 24 h in CFA medium with 100 μM IPTG at 37°C. Cell lysates were assayed by immunoblot analysis using a 1:5000 dilution of anti-PIA antibody. (C) Five independent cultures of wild-type MG1655, the isogenic ΔabgR-ydaL mutant lacking chromosomal mcaS, the chromosomal MG1655 mcaS∷mcaS-3 and MG1655 mcaS∷mcaS-8 mutants, and the isogenic ΔcsrB∷kan mutant were grown in CFA medium for 24 h at 37°C in a 96-well microtiter plate. Biofilm formation was determined by crystal violet staining, measuring absorbance at OD570, and normalizing by OD600. Images in A–C are representative of at least two independent experiments.