Figure 1. Structural and transcriptional analyses of wild-type and aimless1 mutant strains. (A) Schematic illustration of mutagenesis: organization of aim1 and flanking genes on the F. nucleatum ATCC 23726 wild-type and aimless1 mutant chromosome. Arrows indicate PCR primers used for construction and analysis of the aim1 mutant; arrowheads indicate real-time PCR primers; black shaded area indicates intergenic region; bracket indicates location of autotransporter domain. Full-length aim1 (5625 bases) was submitted to GenBank and has been assigned accession number DQ020251. The Fig. is not to scale. (B) PCR analysis confirming insertion into aim1. PCR analysis of chromosomal DNA from ATCC 23726 and the aimless1 mutant. Plasmid pIP-aim1, the construct used to create the inactivation in aim1, was used as the control DNA template. (C) Transcriptional analysis of aim1 and the downstream gene Fn2057. The relative RNA abundance of F. nucleatum wild-type and aimless1 was determined by real-time PCR. Results are expressed as relative means (mutant/wt) ± SEMs (error bars) of triplicate experiments.