Figure 4

oxyR and soxR replacement. (a) Schematic representation of the linear DNA constructed for the oxyR gene replacement. The oligonucleotides used (small arrows) are listed in Table 2. (b) Screening of oxyR A. baumannii mutants generated by gene replacement. The numbers at the top are bacterial colony numbers. WT; Wild-type control showing 1600 bp. Colonies 4 and 7 (lanes 4* and 7*) showing 2275 bp (1600 pb - 258 bp [from oxyR deletion] + 933 bp [from kanamycin insertion]) were sequenced to confirm gene replacement. Lambda DNA-Hin d III and ϕX174 DNA-Hae III Mix (Finnzymes) was used as a size marker (M). (c) Schematic representation of the linear DNA constructed for the soxR gene replacement. The oligonucleotides used (small arrows) are listed in Table 2. (d) Screening of soxR A. baumannii mutants generated by gene replacement. WT: Wild-type control with 1300 bp. Colonies 1, 2, and 3 (lanes 1*, 2*, and 3*) with 2093 bp (1300 bp - 140 bp [from soxR deletion] + 933 bp [from kanamycin insertion]) were sequenced to confirm gene replacement. Lambda DNA-Hin d III and ϕX174 DNA-Hae III Mix (Finnzymes) was used as a size marker (M).