ABSTRACT
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.
Subject(s)
Bacillus subtilis/genetics , Drug Resistance, Microbial , Escherichia coli/genetics , Genetic Vectors , Kanamycin/pharmacology , Base Sequence , Chromosome Mapping , Cloning, Molecular , DNA Restriction Enzymes , Genes, Bacterial , PlasmidsABSTRACT
A hybrid plasmid capable of replication in 2 different genera, Escherichia and Pseudomonas, was constructed. This plasmid DNA can be used as a cloning vector in E. coli and pseudomonades cells. The described hybrid plasmid pLD411 has been constructed on the basis of 2 small E. coli vector R-plasmids used in our laboratory and cryptic plasmid pWW2 or P. putida MT1. Plasmid pLD411 DNA was mapped with restrictases; its biological activity in transformations of different bacterial strains was studied, and the characteristics of transformed cells were also described.