Figure 4.

Characterization of strain MW906. (A) Plasmid yield. Labeling at the top of the panel indicates the combinations of strains and plasmids. For each combination, plasmid DNA was isolated from 2 OD₆₀₀ units of an overnight culture using a standard protocol (for details, see Material and Methods). The marker was the 1 kb Plus DNA Ladder (Life Technologies). The gel used for electrophoresis was composed of 0.8% [w/v] agarose and stained with ethidium bromide. (B) Growth in liquid culture using overnight cultures as an inoculum. The crosses correspond to data-points for MW906 cells containing pGreenII. Compare with the diamonds and squares corresponding to data-points for DH5α cells containing pET28a (Figure 1) and pGreenII (Figure 3), respectively. The inset shows the morphology of MW906 (pGreenII) cells, as imaged in Figure 1, panel A. (C) Restriction enzyme analysis of plasmid from a selection of clones isolated from a culture of MW906 (pMET1-03) following overnight incubation (labeled 1–16). The lane labeled M contains the 1 kb Plus DNA Ladder (Life Technologies). Numbering on the left of the panel indicates the expected sizes of the two fragments produced by EcoRI digestion of pMET1-03. The smaller of the two fragments corresponds to the MET1 cassette destined for plants. The gel used for electrophoresis was composed of 0.8% [w/v] agarose and stained with ethidium bromide.