Figure 4From: Potential complications when developing gene deletion clones in Xylella fastidiosa Mixture of wild-type and mutant Xylella fastidiosa strains after second isolation. The XfΔpilJ mutant confirmed in Figure 3 (isolate 12) was streaked onto periwinkle agar plates amended with kanamycin and the genotype assessed for 32 single colonies. Each number denotes a single colony. A. The pilJE/pilJF (EF) primers amplified a 2030 bp band for non-transformed bacteria and no equivalent bands for the transformed XfΔpilJ mutant strains or the deletion plasmid (P). B. The pilJA/pilJD (AD) primers amplified a 3082 bp band for non-transformed cells and a 2200 bp fragment from the XfΔpilJ strain and the deletion plasmid (P). Wild-type X. fastidiosa DNA (wt) was used as a positive control for the PCR reactions, while primer reaction without template DNA represented by H2O, was used as negative controls for each PCR reaction.Back to article page