Fig. (1) Construction of the plasmid pT7−His−sfGFP−mutA(A) Schematic illustration of the plasmid pT7−His−sfGFP−mutA, with an enlarged part (up), which represents the coding region for the fusion protein sfGFP−mutacin. The most important elements of the plasmid are indicated. These include T7 promoter, His tags, TEV cutting site and the restriction sites used for cloning. Locations of T7 primers (F and R) used for PCR amplification and for sequencing are shown on the plasmid. (B) Agarose gel electrophoresis of PCR products using T7F/T7R primers and the plasmids pT7−His−mutA (lane 1) or pT7−His−sfGFP−mutA (lane 2) as DNA template. (C) DNA amplicons from the previous PCR were digested with NheI and separated by gel electrophoresis. DNA molecular weight marker was loaded in the first lane (M).