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The Streptomyces plasmid SCP2*: its functional analysis and development into useful cloning vectors
Authors:D J Lydiate  F Malpartida  D A Hopwood
Institution:John Innes Institute Colney Lane, Norwich NR4 7UH U.K. Tel. 060352571
Abstract:Detailed restriction maps of the plasmid SCP2* and its deletion derivative pSCP103 were constructed. DNA fragments carrying hygromycin (Hyg), thiostrepton (Thio) or viomycin-resistance (VioR) determinants were inserted into pSCP103, and various segments were deleted from the resulting plasmids. Changes in plasmid phenotypes associated with these insertions and deletions allowed the localisation and characterisation of plasmid replication, stability, transfer and fertility functions. Several useful cloning vectors were constructed. They are able to maintain large (greater than 30 kb) DNA inserts, with stable inheritance at a low copy number (1-2 per chromosome) and without structural rearrangements, in Streptomyces hosts. The vectors have a broad host range in the genus Streptomyces. One of them (pIJ903) is a shuttle vector for Streptomyces and Escherichia coli.
Keywords:Restriction maps  recombinant DNA  hygromycin  thiostrepton and viomycin-resistant derivatives  stable  low-copy-number vectors  Ap  ampicillin  ccc  covalently closed circular  EtBr  ethidium bromide  Hyg  hygromycin  kb  kilobase pairs  MM  minimal medium  Neo  neomyein  resistance  sensitivity  Spc  spectinomycin  Sm  streptomycin  Thio  thiostrepton  Vio  viomycin
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