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Isolation and characterization of pock-forming plasmids for Streptomyces griseus from soil actinomycetes
Authors:T Ohnuki  T Imanaka  S Aiba
Affiliation:Department of Fermentation Technology, Faculty of Engineering, Osaka University, Yamada-oka, Suita-shi, Osaka 565, Japan Tel. 06-877-5111 Ext. 4351
Abstract:Thirty independent actinomycetes strains carrying plasmids were isolated from soil. These plasmids were purified as cccDNA by CsCl-EtBr equilibrium density-gradient centrifugation. Plasmids that induce "pocks", namely formation of circular zones of sporulation-inhibition, were selected by protoplast transformation of streptomycin-producing strain, Streptomyces griseus ATCC10137. Six pock-forming plasmids, pOA7, pOA11, pOA15, pOA23, pOA29 and pOA30, were obtained, and their cleavage maps, transformation frequencies, and copy numbers, as well as their stability, are described.
Keywords:Lethal zygosis  restriction maps  streptomycin  cccDNA  covalently closed circular DNA  EtBr  ethidium bromide  TES  see Materials and Methods, section b
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