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

János Sümegi, Andor Udvardy, Pál Venetianer

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)305-312
Number of pages8
JournalMGG Molecular & General Genetics
Volume151
Issue number3
DOIs
StatePublished - Jan 1 1977

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rRNA Genes
Escherichia coli
Ribosomal RNA
DNA
DNA-Directed RNA Polymerases
Ribosomal DNA
Heparin
Bacterial RNA
DNA-Directed DNA Polymerase
Salts
Molecular Weight
RNA
Weights and Measures
In Vitro Techniques

ASJC Scopus subject areas

  • Genetics

Cite this

In vitro transcription of the ribosomal RNA genes of E. coli DNA. / Sümegi, János; Udvardy, Andor; Venetianer, Pál.

In: MGG Molecular & General Genetics, Vol. 151, No. 3, 01.01.1977, p. 305-312.

Research output: Contribution to journalArticle

Sümegi, János ; Udvardy, Andor ; Venetianer, Pál. / In vitro transcription of the ribosomal RNA genes of E. coli DNA. In: MGG Molecular & General Genetics. 1977 ; Vol. 151, No. 3. pp. 305-312.
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