Differential Regulation of Escherichia coli fim Genes following Binding to Mannose Receptors

Figure 4

Quantitative determination of the fimS invertible element orientation of E. coli cells with a plasmid that has the FimH protein represented as wild type (WT), Null, Q133K, or N46A mixed with mannose-coated Sepharose beads (+) for 0 h, 4 h, or 24 h. The PCR analysis was performed with twofold dilutions of chromosomal DNA isolated from the E. coli cells using the INV and FIMA primers to amplify Phase-ON-oriented DNA (450 bp product) or FIME and INV primers to amplify Phase-OFF-oriented DNA (750 bp product). All PCR products were electrophoresed on 1.5% agarose gels. The following dilutions of DNA were used: undiluted (lane 1), 1/2 (lane 2), 1/4 (lane 3), 1/8 (lane 4), 1/16 (lane 5), 1/32 (lane 6), 1/64 (lane 7), 1/128 (lane 8), 1/256 (lane 9), and 1/512 (lane 10). The data represent at least three separate runs.