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. 2011 Nov 24;6(5):975–983. doi: 10.1038/ismej.2011.166

Figure 6.

Figure 6

Binding of GSU0514 to the sucC–sucD promoter. (a) GSU0514-binding site. Footprint assays were carried out with GSU0514 and the upstream region of the sucC–sucD operon. DNA-binding reactions were conducted with GSU0514 (lane 1, 0 pmol; lane 2, 1.3 pmol and lane 3, 2.5 pmol) and probes (0.2 pmol). G, A, T and C represent sequence ladders. The region protected from or modified by DNaseI in the presence of GSU0514 is indicated by a vertical bar. (b) Promoter region of the sucC–sucD operon. The GSU0514-binding site identified by the footprint assay (a) is indicated by bold letters with a box. Two putative transcription initiation sites determined by primer extension assays (data not shown) are indicated by +1. Putative −35 and −10 promoter elements are indicated by −35 and −10, respectively. Putative ribosome-binding site is indicated by RBS. Translation initiation codon is indicated by Met. (c) DNA-binding assay. DNA fragments containing promoter regions of the sucC–sucD operon were tested for DNA-binding activity of GSU0514. The promoter regions in the DNA fragments (1∼3) used as a probe are shown below the diagram of the sucC–sucD operon. Arrows indicate putative transcription initiation sites. Black box represents the GSU0514-binding site identified by the footprint assay (a).