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An asd-deleted (Δasd) mutant of Salmonella enterica serovar Pullorum (SP) was constructed using an improved method of gene knockout by combining the π-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 Δasd mutant was further defined. The improved method was simpler and more effective than previously reported conventional methods

An Improved Method to Knock Out theasdGene ofSalmonella entericaSerovar Pullorum