Molecular cloning of genetically active fragments of Bacillus DNA in Bacillus subtilis and properties of the vector plasmid pUBi 1 0 ( genetic complementation / tryptophan / EcoRl endonuclease ) KATHLEEN
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چکیده
ABSTRACr Plasmid pUB10 (-2.8 X 106 daltons), originally detected in Staphylococcus aureus, specifies resistance to neomycin and has been transformed into Bacillus subtilis 168. In B. subtifis, pUBllO is stably maintained at about 50 copies per chromosome and renders the host resistant to neomycin sulfate at 5 jg/ml. pUBlIO isolated from B. subtifis transforms Rec+ and recE4-containing strains of B. subtifis at frequencies >103 transformants per jsg of plasmid. pUBlIO was transferred by PBS1 or SPIO transduction from B. subtifis to strains of B. pumilus and B. licheniformis. pUBl10 is compatible with each of four previously describe Bacillus plasmids, including pPL576, pPL1O, pPL706S, and pPL2. pUlBIlO contains a single EcoRI-sensitive site and was used as vector to clone DNA fragments that complement the trpC2 mutation in B. subtilis 168 from EcoRl digests of the chromosome DNA isolated from B. pumilus strains NRRL B-3275 and NRSS76, B. Iicheniformis strains 9945A and 749C, and B. subtilis 168. Genetic and physical properties of each of the constructed Trp derivatives of pUBI1O are described.
منابع مشابه
Recombination between compatible plasmids containing homologous segments requires the Bacillus subtilis recE gene product.
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