Figure 3.

Southern blot analysis confirms the deletion of the carRP gene and excision of the pyrG marker gene. Southern blotting was performed to ensure the deletion of the carRP gene and excision of the pyrG marker. Genomic DNA of the wild-type strain was digested with BclI and produced a 5011-bp fragment that was recognized by the probe. A double digestion with BclI and EcoRI resulted in a 1322-bp fragment that was recognized by the probe. The digestion of the genomic DNA of the carRPΔ::pyrG-dpl237 mutant with BclI produced a 2501-bp fragment that was recognized by the probe. The double digestion with the enzymes BclI and EcoRI produced an 872-bp fragment recognized by the probe. Finally, the carRPΔ::dpl237 mutant underwent the same digestions, where a 3539-bp fragment was produced after BclI enzyme digestion, and an 870-bp fragment was produced after BclI and EcoRI double digestion. Both fragments were detected during our hybridization procedure by our probe. Gene size is not to scale.