Transconjugation plasmid for mutating Pasteurellaceae, useful for producing live vaccine strains of the pig pathogen Actinobacillus pleuropneumoniae

摘要

A transconjugation plasmid (A) for introducing unmarked mutations into the chromosome of bacteria Pasteurellaceae comprising: (a) the mobilization region mobRP4; (b) a polylinker; (c) a replicon of the ColE1 type; (d) a resistance determinant; and (e) a transcription fusion between the sacB gene of Bacillus subtilis and the omlA promoter of Actinobacillus pleuropneumoniae, is new. Independent claims are also included for the following: (1) a method (M1) for the mutagenesis of Pasteurellaceae by transconjugation with a plasmid (A') similar to (A) that includes an exogenous DNA segment (I) in the polylinker; Pasteurellaceae transformed with (A'); (2) a host cell stably transformed with (A) or (A'); and (3) a kit for introducing unmarked deletions into the chromosomes of Pasteurellaceae containing (A) or (A').