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A vector for recombinant DNA in Staphylococcus aureus
Authors:Sven Löfdahl  Jan-Eric Sjöström  Lenhart Philipson
Institution:Department of Microbiology, Biomedical Center, Box 581, S-751 23 Uppsala, Sweden
Abstract:Staphylococcal plasmids pS194 and pSC194 which confer streptomycin and streptomycin-chloramphenicol resistance respectively have been used as vectors for construction of recombinant DNA, since they each carry one single recipient site for endonuclease EcoRI. Hybrid DNA does not express streptomycin resistance, a marker which is present in both vectors, presumably because the marker gene is cleaved by EcoRI. A chloramphenicol marker present in pSC194 was used for positive hybrid selection. Hybrid plasmids generated by joining pSC194 with one or more of the four EcoRI fragments of the large (18.1-10(6) daltons) staphylococcal plasmid pI258 were constructed and permitted us to develop a physical map for pI258.
Keywords:Restriction enzymes  plasmid  streptomycin and chloramphenicol resistance  gene maps  ccc-DNA  covalently closed circular DNA  SDS  sodium dodecyl sulphate  TE  Tris-EDTA  TEB  Tris-EDTA-borate  TSA  Trypticase Soy Agar
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