中国生物工程杂志

Mutant construction is a prerequisite work in bacterial gene function studies. In this report, we constructed a new suicide plasmid to construct Brucella mutant. A kanamycine gene was inserted at the multi-cloning site of pUC19, and on either side of the resistance gene, several enzyme sites were inserted, resulting in the new plasmid pUC19K. Using this plasmid, we constructed replacement mutant of Brucella omp25 gene. As the results showed, the mutant could be generated by only one round of selection with high efficiency. Therefore, the construction and application of pUC19K provides a new suicide plasmid for Brucella mutant construction and would accelerate gene function analysis of Brucella.