An Improved Method to Knock Out the asd Gene of Salmonella enterica Serovar Pullorum

An asd-deleted (Deltaasd) mutant of Salmonella enterica serovar Pullorum (SP) was constructed using an improved method of gene knockout by combining the pi-suicide plasmid system with the Red Disruption system. The asd gene was efficiently knocked out by the recombinant suicide vector, which replaced the asd gene with the CmR gene. Based on the balanced lethal host-vector system, the phenotype of the Deltaasd mutant was further defined. The improved method was simpler and more effective than previously reported conventional methods.