Molecular and General Genetics MGG

, Volume 143, Issue 3, pp 291–295 | Cite as

Quantitation of RNA polymerase subunits in Escherichia coli during exponential growth and after bacteriophage T4 infection

  • Frode Engbaek
  • Carol Gross
  • Richard R. Burgess
Article
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Summary

A method was developed to measure the amounts of RNA polymerase subunits, α, β, β′ and σ in crude extracts of Escherichia coli. The proteins were labelled by growing the cells in 35S-sulphate containing media. For measuring β and β′, the cell lysate was electrophoresed on 6% polyacrylamide gels containing SDS and the β and β′ bands cut out and counted. For measuring α and σ, the cell lysate was co-electrophoresed with dansylated RNA polymerase on 8% polyacrylamide gels containing SDS. The fluorescent bands were cut out, the proteins eluted, and the α and σ subunits further purified on polyacrylamide gels containing 8 molar urea.

The results are: (1) β′ is the subunit of the core RNA polymerase which is present in limiting amount. (2) The core enzyme, as measured by β′, constitutes a constant fraction of total cellular protein (0.9%), independent of the bacterial growth rate. (3) The α subunit is made in excess and is probably regulated independently. (4) The σ subunit is present in 0.3–0.4 times the amount of the core enzyme. (5) All four subunits are fully conserved after bacteriophage T4 infection.

Keywords

Escherichia Coli Urea Polyacrylamide Crude Extract Bacterial Growth 
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 1976

Authors and Affiliations

  • Frode Engbaek
    • 1
  • Carol Gross
    • 1
  • Richard R. Burgess
    • 1
  1. 1.McArdle Laboratory for Cancer ResearchUniversity of WisconsinMadison

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