Figure 2

Verification of the restriction maps of GS_43D16. A, KpnI and NotI map of the assembled GS_43D16 sequence. B, KpnI and NotI double digestion of selected GS_43D16 clones carrying the EZ::TN <NotI/KAN-3> transposon insertions. Eight fragments were expected from KpnI and NotI digestion of GS_43D16 carrying no transposons (A). Only five fragments were observed, because some of the fragments had similar mobilities in the gel. Some of these fragments were resolved after transposon insertion. A close relationship was observed between the restriction fragment sizes determined by gel electrophoresis and that by sequence data and location of transposon insertions (Table 1). m1, λHindIII ladders, m2, 1 kb DNA ladder (New England Biolabs Inc., Beverly, MA). C, SalI-NotI map of the assembled GS_43D16 sequence. D, SalI and NotI digestion of GS_43D16. Eight fragments were expected from the double digestion of GS_43D16 (Figure 2C). Six fragments were resolved from the digestion of the clone (43 in 2D). 7.9 kb and 7.11 kb fragments were not resolved (Fragment IV, twice the intensity of either Fragment III or Fragment V) and 0.6 kb SalI-NotI fragment is not included in 2D.