Fig. 3
Southern blot analysis of strains of B. subtilis and G. kaustophilus. A Schematic organization of the relevant chromosomal loci of B. subtilis YNB213 and G. kaustophilus MK244 and MR01. Chromosomal segments corresponding to B. subtilis and G. kaustophilus are shown as thinner and thicker lines, respectively. The ΔiolQGK::kan cassette set in the chromosome of YNB213 is shown at the top in brackets. ClaI and SacII sites are indicated with a scale underneath representing the relative distance (bp) from the most upstream ClaI site. The kan and iolQGK genes and oriT are shown as arrowheads. The stretch of probe set within kan for the Southern blot analysis is indicated as a solid rectangle. B Results of the Southern blot analysis using the RNA probe to detect kan. Chromosomal DNAs of MK244, YNB213, and MR01 were digested by ClaI and/or SacII as indicated and subjected to the analysis. No signal was detected in MK244, since it does not have kan. In YNB213 and MR01, a single band specific to each was detected. Only for the digestion by ClaI, the length of the DNA fragments containing kan varies due to the difference in the restriction sites. The experiments were repeated at least three times with similar results, and the representative data were shown