Mini-Tn10 transposon derivatives for insertion mutagenesis and gene delivery into the chromosome of gram-negative bacteria.

Four mini-Tn10 derivatives bearing the nptII, cat, aacC1 and tet genes along with MCS of the pBluescriptII plasmid were constructed. These derivatives are embedded into the gamma-ori R6K-based suicide plasmid pBSL177, which can be mobilized into a broad range of hosts by the RP4 plasmid. The relatively small size and the presence of the mutant ATS Tn10 transposase gene, fused to Ptac promoter, as well as the versatile MCS of the pBluescriptII plasmid make these vectors a reliable tool for insertion mutagenesis and chromosomal insertion of the cloned DNA fragments.