Versatile Escherichia coli-Bacillus shuttle vectors derived from runaway replication plasmids related to CloDF13

Abstract

Versatile cloning vectors were constructed employing a runaway replication mutant of the bacteriocinogenic plasmid CloDF13. These vectors can, under conditions where protein synthesis is not inhibited, be amplified in Escherichia coli to high levels by elevating the temperature and are therefore useful for the production of large quantities of DNA and protein. Since the constructed shuttle vectors, which harbour at least six unique restriction endonucleases sites, replicate in E. coli, Enterobacter cloacae, Staphylococcus aureus and a variety of Bacilli, they are applicable for the genetic engineering of both gramnegative and gram-positive bacteria.