FIG. 1.

PFGE and Southern hybridization analysis of S. roseosporus mutants containing Tn5099 insertions. Southern hybridizations were carried out with a gel-purified 4-kb HindIII fragment from pCZA213 containing Tn5099 as the probe. (A) Contour-clamped homogeneous electric field (CHEF) gel and Southern blot of S. roseosporus DNA cleaved with DraI. A 1% FastLane agarose gel in 0.25× Tris-borate-EDTA buffer (TBE) was run at 6 V/cm and 14°C at an angle of 120°C with a 30- to 90-s pulse (linear over 16 h). (B) CHEF gel and Southern blot of S. roseosporus DNA cleaved with AsnI. A 1% FastLane agarose gel in 0.1× TBE was run at 6 V/cm and 16°C at an angle of 120°C with a 2- to 30-s pulse (linear over 15 h). Note that strain A21978.65 lacks the 180-kb AsnI-K₂ fragment present in the other strains, which are derived from A21978.6 (20). (C) CHEF gel and Southern blot of S. roseosporus DNA cleaved with AsnI. A 1% FastLane agarose gel in 0.25× TBE was run at 6 V/cm and 14°C at an angle of 120°C with a 60- to 110-s pulse (linear over 24 h).