Fig 2.

Construction of a fliA deletion mutant. (A) Schematic diagram showing the mutagenesis strategy employed to construct the fliA deletion mutant of C. ljungdahlii. Homologous recombination via a suicide vector, pBuTU-ΔfliA-ermC, resulted in three possible genotypes: double crossover, single crossover at the 5′ homologous region, and single crossover at the 3′ homologous region. (B) PCR analysis of representative fliA deletion mutants. Genotypes were verified by PCR amplification with distal primers indicated by arrows in panel A. The PCR product from the wild-type locus (lane 1) is smaller than that from the mutant locus (lanes 2 and 3) because the replaced region in the fliA gene is smaller than the ermC gene. Lane L, 1-kb DNA ladders (New England BioLabs); lane 1, wild type; lane 2, double-crossover fliA mutant; lane 3, single-crossover fliA mutant.