Open AccessElectron microscopic analysis of transcription: mapping of initiation sites and direction of transcription.
Author(s) -
Christine Brack
Publication year - 1979
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.76.7.3164
Subject(s) - transcription (linguistics) , general transcription factor , rna polymerase , promoter , transcription factor ii d , microbiology and biotechnology , rna polymerase ii , biology , transcription bubble , abortive initiation , rna , genetics , gene expression , gene , philosophy , linguistics
An electron microscope technique is described that allows rapid characterization of transcription in vitro. DNA is transcribed with Escherichia coli RNA polymerase in vitro, and the RNA is hybridized to its template. Measurement of the resulting transcription R-loop molecules allows accurate mapping of transcription initiation sites (promoter sites) and analysis of the direction and rate of transcription and the level of transcription from each initiation site. The two major early promoters pR and pL of bacteriophage lambda have been mapped within 0.1-0.3 map units of the known positions and three additional sites have been confirmed. Six transcription initiation sites have been preliminarily mapped on plasmid pSF2124 DNA.