FIG. 3.

Strategy for mutagenesis and construction of plasmids expressing GtfB and -C with insertion, deletion, and single amino acid mutations. Black bars represent gtfB and gtfC genes. pYNB13-D6 and -D201 were generated by cloning of the PCR-derived fragment containing in-frame deletion of 14 aa. pNH3-D1 was generated by a similar procedure. pAlter1-EB was generated by cloning of EcoRI-BamHI fragment from gtfC into identical multiple cloning sites in plasmid pAlter1, which carries two antibiotic resistance markers, ampicillin and tetracycline. nt, nucleotides.