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  • Title: Construction and propagation of deletion derivatives of staphylococcal tetracycline-resistance plasmid pTP-5 in Bacillus subtilis.
    Author: Noguchi N, Shishido K, Ando T, Kono M.
    Journal: Gene; 1983; 21(1-2):105-10. PubMed ID: 6301940.
    Abstract:
    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.
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