Bacterial vectors which confer resistance to kanamycin

On the base of plasmid pLD720 (a deletion derivative of the cosmid vector pHC79) a number of hybrid plasmids which confer in Escherichia coli cells the kanamycin resistance was constructed. All hybrid plasmids contain the promoterless part of kanamycin resistance gene (which codes for aminoglycoside 3'-phosphotransferase II) from transposon Tn5. The Km gene expression is driven by a promoters situated on pLD720. The hybrid plasmids pLD723, pLD724 and pLD728 contain a complete DNA sequences of plasmids pC194 or pE194 from Staphylococcus aureus that permits them to replicate into Bacillus subtilis as well. However, no expression of the Km gene in Bacillus subtilis was observed. There is a unical Bgl II site on pLD728 is front of the beginning of a Km gene structural part. This property of pLD728 may be useful when cloning in this plasmid a promoter sequences of different species.