FIG. 3.

Restriction maps of constructions used to inactivate CT1987 and CT0967 and confirmation of segregated mutants. (A) Restriction maps and primer locations of constructions used to inactivate CT1987 and CT0967. (B) Electrophoretic analysis of PCR products to evaluate putative mutants. The CT1987 and CT0967 loci were amplified by PCR with the primers indicated in Table 1. The DNA templates were derived from wild-type C. tepidum (lanes 2 and 4), from a CT1987::aadA transformant (lane 3), and from a CT0967::aadA transformant (lane 5). The amplicons from the two transformants are 0.9 kb larger than the corresponding amplicon from the wild type. This demonstrates that wild-type and mutant alleles had segregated fully in both transformants. Lanes 1 and 6 contain DNA size markers (Ladder I; GeneChoice, Frederick, MD).