Figure 10
From: Characterisation of a natural variant of the γ-butyrolactone signalling receptor
Gel retardation assay using ScbR M145 and ScbR M600 from E. coli supplemented with SCB1. DNA and γ-butyrolactone binding abilities of ScbRM145 and ScbRM600 from E. coli cell-free extracts (CE) were tested using a DIG-labelled scbR promoter DNA fragment and the S. coelicolor γy-butyrolactone SCB1 in a gel retardation assay. All samples contained the labelled DNA probe. Sample two contained CE of E. coli/pIJ2925 (“E. coli CE”), samples 3–6 and 7–10 of E. coli JM101/pIJ6120 and pTE58 (“ScbRM145” and “ScbRM600”). To samples 4, 5, 8 and 9 SCB1 dissolved in methanol was added in high (63 ng) and low (8 ng) amounts. Samples 6 and 10 were supplemented with same volumes of pure methanol. The scbR promoter DNA fragment and DNA/protein complexes formed with ScbRM145/M600 are indicated by arrows. DNA binding abilities of the two variants of ScbR were shown to be the same in the absence and the presence of the γ-butyrolactone.