Formation of chimeric DNA primer extension products by template switching onto an annealed downstream oligonucleotide

Proc Natl Acad Sci U S A. 1996 Apr 2;93(7):2969-74. doi: 10.1073/pnas.93.7.2969.

Abstract

Oligodeoxynucleotide sequences are described that anneal to a template downstream of a priming site. During polymerase-catalyzed extension of the primer, the extending primer shifts from the original template to a segment of the annealed oligonucleotide that acts as an alternative template. The resulting chimeric extended primer has one segment that is complementary to the template and a second segment that is complementary to the oligonucleotide. The influence of the sequence elements of the oligonucleotide and the reaction conditions on template switching have been explored. The sequence requirements for template switching are compared to those for transposon excision.

Publication types

  • Comparative Study

MeSH terms

  • Base Sequence
  • Binding Sites
  • DNA Primers*
  • DNA, Single-Stranded
  • DNA-Directed DNA Polymerase / metabolism*
  • Escherichia coli / genetics
  • Genes, Bacterial
  • Models, Structural
  • Molecular Sequence Data
  • Nucleic Acid Conformation*
  • Polymerase Chain Reaction
  • Structure-Activity Relationship
  • Substrate Specificity
  • Templates, Genetic

Substances

  • DNA Primers
  • DNA, Single-Stranded
  • DNA-Directed DNA Polymerase