Supporting information for Liu et al. (2003) Proc. Natl. Acad. Sci. USA, 10.1073/pnas.0630387100
Fig. 7.
Resistance phenotype, construction, and genomic structure for the DR0070 knockout mutant. (A) Survival of DEIRA strains MD891 (DR0070 disruption) and MD68 (DEIRA wild-type strain R1/pMD68) following γ-irradiation. Cells were grown to the early plateau phase and irradiated on ice to the indicated doses. Aliquots were withdrawn at intervals, diluted, and plated on TGY agar that was supplemented with Km. Squares, MD68; circles, MD891. Values are the means ± SD of three independent experiments. (B) Predicted DNA band sizes of wild-type DR0070 arising from cleavage with the indicated restriction endonucleases. ATG and TGA, DR0070 initiation and termination codons, respectively. (C) Predicted DNA band sizes of disrupted DR0070 after integration of pCR2.1, arising from cleavage with the indicated restriction endonucleases. ATG and TGA, as in B. (D) MD891 was subjected to a detailed mapping of the DR0070 integration site by using restriction enzymes indicated in B and C, Southern blotting, and probing with a 575-bp radiolabeled DR0070 fragment shown in B. wt, wild-type (DEIRA); m, mutant (MD891). Molecular size standards (kbp): 1, DNA Ladder Mix (Fermentas, Hanover, MD); 2, 200-bp DNA Step Ladder (Promega); 3, 2-Log DNA Ladder (New England Biolabs).