Molecular and General Genetics MGG

, Volume 151, Issue 3, pp 305–312 | Cite as

In vitro transcription of the ribosomal RNA genes of E. coli DNA

  • János Sümegi
  • Andor Udvardy
  • Pál Venetianer
Article
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Summary

Bacterial ribosomal RNA synthesis was studied in an in vitro system in which the presence of heparin prevented reinitiation of transcription. The number of heparin-resistant binary complexes of RNA-polymerase and E. coli DNA depended strongly on the quality of the template. High-molecular weight DNA was a much superior template than DNA prepared by conventional techniques. Using this highmolecular weight DNA as template the amount of ribosomal RNA synthetized in one round of transcription was found to be 4–5 fold higher than the amount of rDNA present. Controls have shown that the transcription probably started at the proper initiation sites and no significant read-through form distant promoters contributed to this effect. If the binary polymerase-DNA complexes were dissociated in the presence of 0.5 M KC1 prior to transcription all RNA synthesis was strongly reduced but the proportion of rRNA increased in the transcript. However, in this case the amount of rRNA did not exceed the amount of rDNA. We propose that the promoters of the rRNA genes are complex structures, able to store 4–5 molecules of RNA polymerase and of these several polymerase only one is bound in an extremely salt-resistent form.

Keywords

Heparin Initiation Site Conventional Technique Binary Complex Distant Promoter 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
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Copyright information

© Springer-Verlag 1977

Authors and Affiliations

  • János Sümegi
    • 1
  • Andor Udvardy
    • 1
  • Pál Venetianer
    • 1
  1. 1.Institute of Biochemistry, Biological Research CenterHungarian Academy of SciencesSzegedHungary

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