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Figure 8 | BMC Research Notes

Figure 8

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

Figure 8

Gel retardation assay and Western analysis using S. coelicolor cell-free extracts. Cell-free extracts (CE) of S. coelicolor LW34 (scbRM145) and LW33 (scbRM600)obtained at four time points (tp 1–4) during different phases of growth indicated with eT, mT, lT and S (e arly, m id, l ate t ransition, and s tationary phase; also see Table 2, GC 1) were used to determine presence and DNA binding ability of ScbRM145/M600. CE from E. coli JM101/pIJ6120 harbouring ScbRM145 was used as a positive control (indicated with a “+”). A. DNA binding abilities of ScbRM145 and ScbRM600 from S. coelicolor CE were tested using a DIG-labelled scbR promoter DNA fragment by gel retardation analysis. The scbR promoter DNA fragment and DNA/protein complexes are indicated by arrows. The DNA probe alone was used as a negative control (indicated with a “-“). B. Western analysis of ScbR. ScbRM145 (+, LW34) and ScbRM600 (LW33) was detected in similar amounts at corresponding time points in the two S. coelicolor strains. ScbR signals are indicated by an arrow.

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