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Figure 11 | BMC Research Notes

Figure 11

From: Characterisation of a natural variant of the γ-butyrolactone signalling receptor

Figure 11

Gel retardation assay and Western analysis using ScbR M145 and ScbR M600 from E. coli after freezing and thawing. A. DNA binding abilities of ScbRM145 and ScbRM600 from E. coli cell-free extracts (CE) were tested using a DIG-labelled scbR promoter DNA fragment in a gel retardation assay. All samples contained the labelled DNA probe. Samples 2–5 and 6–9 contained CE of E. coli JM101/pIJ6120 and pTE58 (“ScbRM145” and “ScbRM600”), respectively, frozen and thawed up to three times before added to the DNA probe. The scbR promoter DNA fragment and DNA/protein complexes formed with ScbRM145/M600 are indicated by arrows. In contrast to ScbRM145, DNA binding of ScbRM600 was shown to be unstable under the conditions tested. B. Western analysis of ScbR. ScbRM145 was detected with a positive control sample of recombinant ScbRM145 (“rScbRM145”, lane 1) and in comparable amounts in all gel retardation assay samples harbouring ScbRM145 or ScbRM600 as described in A (lanes 2–9). ScbR bands are indicated by an arrow.

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