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Isolation and characterization of naturally occurring hairpin structures in single-stranded DNA of coliphage M13
Authors:S K Niyogi  S Mitra
Institution:Biology Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee 37830 USA
Abstract:With precise conditions of digestion with single-strand-specific nucleases, namely, endonuclease S1 of Aspergillus oryzae and exonuclease I of Escherichia coli, nuclease-resistant DNA cores can be obtained reproducibly from single-stranded M13 DNA. The DNA cores are composed almost exclusively of two sizes (60 and 44 nucleotides long). These have high (G + C)-contents relative to that of intact M13 DNA, and arise from restricted regions of the M13 genome. The resistance of these fragments to single-strand-specific nucleases and their nondenaturability strongly suggest the presence of double-stranded segments in these core pieces. That the core pieces are only partially double-stranded is shown by their lack of complete base complementarity and their pattern of elution from hydroxyapatite.
Keywords:ss  single-stranded  ds  double-stranded  RF DNA  replicative form DNA  Haemophilus influenzae restriction endonuclease  EGTA  ethylenebis(oxyethylene nitrilo)tetraacetate  TCA  trichloroacetic acid  SDS  sodium dodecyl sulfate
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