A Restriction Enzyme Cleavage Map of Tn5 and Location of a Region Encoding Neomycin Resistance

Mol Gen Genet. 1979;177(1):65-72. doi: 10.1007/BF00267254.

Abstract

This paper reports a cleavage site map of Tn5 for restriction enzymes BamHI, Bg/I, Bg/II, Hind II, HindIII, HpaI, Sa/I, Aval, SmaI, XhoI, PstI, PvuII, HaeII and HaeIII that was determined by the analysis of restriction enzyme cleavage patterns of ColEl, two independent ColEl::Tn5 plasmids, and a ColEl::Tn5 deletion derivative. Ba/I, EcoRI, KpnI, and PvuI do not cleave Tn5. Construction and analysis of in vitro-generated deletions of a ColEl::Tn5 plasmid limit the sequences encoding neomycin resistance to a 1500-base-pair-long segment of Tn5. Insertion of DNA at a Bg/II site within this segment results in loss of the neomycin resistance phenotype. Since this Bg/II site lies in an inverted repeat region, sequences within this repeat seem to be involved in the expression of neomycin resistance.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Base Sequence
  • Chromosome Mapping
  • DNA Restriction Enzymes*
  • DNA Transposable Elements*
  • Drug Resistance, Microbial
  • Escherichia coli / drug effects
  • Escherichia coli / genetics*
  • Genetic Code
  • Mutation
  • Neomycin / pharmacology*
  • Phenotype

Substances

  • DNA Transposable Elements
  • DNA Restriction Enzymes
  • Neomycin