Production of double-stranded RNA during synthesis of bromouracil-substituted RNA by transcription with T7 RNA polymerase

FEBS Lett. 1994 Sep 5;351(2):253-6. doi: 10.1016/0014-5793(94)00870-1.

Abstract

Using T7 RNA polymerase we synthesized a short oligoribonucleotide containing bromouracil by in vitro transcription of a synthetic DNA template. Whereas the major transcript obtained had the expected size and was apparently homogeneous on a sequencing gel, additional analysis revealed the presence of double-stranded RNA in this preparation. As this was not observed when the same template was transcribed in the presence of uracil, we hypothesize that bromouracil promoted the apparition of double-stranded 'parasitic' RNA presumably by favouring priming for the RNA-dependent RNA synthesis of the T7 RNA polymerase or by facilitating an end-to-end copy mechanism.

MeSH terms

  • Base Composition
  • Base Sequence
  • Bromouracil / metabolism*
  • DNA-Directed RNA Polymerases / metabolism*
  • Models, Genetic
  • Molecular Sequence Data
  • Nucleic Acid Denaturation
  • Oligoribonucleotides / biosynthesis*
  • RNA, Double-Stranded / biosynthesis*
  • Transcription, Genetic*
  • Viral Proteins

Substances

  • Oligoribonucleotides
  • RNA, Double-Stranded
  • Viral Proteins
  • Bromouracil
  • bacteriophage T7 RNA polymerase
  • DNA-Directed RNA Polymerases