Fig. 4

PCR validation of DFCT16, an rsbv1 and incA dual-insertion mutant created using the GII(bla) and GII(aadA) cassettes. DFCT13 EBs were transformed with pDFTT3aadA and serially passaged with spectinomycin selection. Mutants were plaque purified and a single plaque was expanded for PCR and phenotype analysis (Figs. 5, 6). The incA and rsbV1 insertion locus maps are shown in panel a. Expected PCR product sizes are listed in panel b and the primers used along with reaction notes are listed in Table 1. PCR products were separated and visualized as described for Fig. 2. PCR results are shown for the wild type strain and DFCT16 in panels c and d, respectively