Production of double-stranded RNA during synthesis of bromouracil-substituted RNA by transcription with T7 RNA polymerase.
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.
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Laboratoire de Biophysique Moléculaire, INSERM U386, Université de Bordeaux II, France.
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D001482:Base Composition; D001483:Base Sequence; D001976:Bromouracil / Q000378:metabolism*; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D008957:Models, Genetic; D008969:Molecular Sequence Data; D009691:Nucleic Acid Denaturation; D009843:Oligoribonucleotides / Q000096:biosynthesis*; D012330:RNA, Double-Stranded / Q000096:biosynthesis*; D014158:Transcription, Genetic; D014764:Viral Proteins
D009843:Oligoribonucleotides; D012330:RNA, Double-Stranded; D014764:Viral Proteins; D001976:Bromouracil; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases