Open AccessTermination of transcription by bacteriophage T3 RNA polymerase: homogeneous 3'-terminal oligonucleotide sequence of in vitro T3 RNA polymerase transcripts.
Author(s) -
Hemanta K. Majumder,
Umadas Maitra,
Martin Rosenberg
Publication year - 1979
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.76.10.5110
Subject(s) - rna , oligonucleotide , microbiology and biotechnology , biology , nuclease protection assay , transcription (linguistics) , polymerase , termination factor , rna polymerase ii , rnase p , rna dependent rna polymerase , rna polymerase , small nuclear rna , gel electrophoresis , t7 rna polymerase , dna , biochemistry , bacteriophage , gene expression , gene , promoter , linguistics , philosophy , escherichia coli
RNA was synthesized in vitro from a T3 DNA template by T3 RNA polymerase and subsequently separated into seven discrete size classes (molecular weights ranging between 0.21 x 10(6) and 6.2 x 10(6)) by electrophoresis in polyacrylamide slab gels. RNase T1-generated 3'-terminal oligonucleotide fragments were then selectively isolated from either the unfractionated total RNA or the gel-purified specific transcripts by chromatography on columns of dihydroxyboryl-cellulose. Sequence analysis of these oligonucleotide products indicated that the unfractionated transcripts as well as all the individual major RNA species examined had a unique sequence, (Gp)UpUpUpUpUpGOH, at their 3' termini. The specificity of this sequence, as well as the total lack of any sequence heterogeneity at the ends of these transcripts, indicates a high degree of specificity of termination during transcription in this system.