Construction and propagation of deletion derivatives of staphylococcal tetracycline-resistance plasmid pTP-5 in Bacillus subtilis.
Tetracycline-resistance (TCr) plasmid pTP-5 (4.46 kb) from Staphylococcus aureus was cleaved with HindIII into three fragments: A (2.35 kb), B (1.57 kb) and C (0.54 kb). A deletion plasmid (3.92 kb) lacking HindIII-C fragment was obtained, and was designated pNS1. This plasmid retained the TCr phenotype and the ability to replicate autonomously in Bacillus subtilis. A restriction endonuclease cleavage map of pNS1 was constructed. Attempts to construct smaller plasmids by digesting pNS1 with BAL31 nuclease led to production of a set of deletion derivatives whose molecular sizes range from 3.75 to 2.77 kb. Through analyses of these derivatives, the regions essential for autonomous replication and expression of TCr were deduced.
D001412:Bacillus subtilis / Q000187:drug effects / Q000235:genetics*; D002872:Chromosome Deletion; D002876:Chromosomes, Bacterial; D004262:DNA Restriction Enzymes; D004352:Drug Resistance, Microbial; D010641:Phenotype; D010957:Plasmids; D013211:Staphylococcus aureus / Q000187:drug effects / Q000235:genetics*; D013752:Tetracycline / Q000494:pharmacology*
D004262:DNA Restriction Enzymes; D013752:Tetracycline