Skip to main content
SpringerLink
Log in
Book cover

New Protein Techniques pp 135–147Cite as

Purification of DNA-Dependent RNA Polymerase from Eubacteria

  • Protocol

  • 1521 Accesses

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 3))

Abstract

In eubacteria, the structure of DNA-dependent RNA polymerase is conserved; the enzyme is composed of at least four different subunits: beta (β), beta-prime (β′), and alpha (α), which form the core (ββ′α2), and sigma (σ), which is required for efficient promoter selection (1–3).

This is a preview of subscription content, log in via an institution.

Protocol
USD   49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   109.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Burgess, R.R. (1976) Purification and Physical Properties of E. coli RNA Polymerase, in RNA Polymerase (Losick, R. and Chamberlin, M., eds.) Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.

    Google Scholar 

  2. Fukuda, R., Ishihams, A., Saitoh, T., and Taketo, M. (1977) Comparative studies of RNA polymerase subunits from various bacteria. Mol. Gen. Genet. 154, 135–144.

    Article  PubMed  CAS  Google Scholar 

  3. Wiggs, J.L., Buch, J.W., and Chamberlin, M.J. (1979) Utilization of promoter and terminator sites on bacteriophage T7 DNA by RNA polymerases from a variety of bacterial orders. Cell 16, 97–109.

    Article  PubMed  CAS  Google Scholar 

  4. Doi, R.H. (1982) RNA Polymerase of Bacillus subtilis, in The Molecular Biology of the Bacilli, vol. 1, Bacillus subtilis (Dubnau, D.D., ed.) Academic, New York.

    Google Scholar 

  5. Sternbach, H., Engelhardt, R., and Lezius, A.G. (1975) Rapid isolation of highly active RNA polymerase from Escherichia coli and its subunits by matrix-bound Heparin. Eur. J. Biochem. 60, 51–55.

    Article  PubMed  CAS  Google Scholar 

  6. Grossman, A.D., Erickson, J.W., and Gross, C.A. (1984) The htp R gene product of E. coli is a sigma factor for heat shock promoters. Cell 38, 383–390.

    Article  PubMed  CAS  Google Scholar 

  7. Westpheling, J., Ranes, R., and Losick, R. (1985) RNA polymerase heterogeneity in Streptomyces coelicolor. Nature 313, 22–27.

    Article  PubMed  CAS  Google Scholar 

  8. Price, C.W. and Doi, R.H. (1985) Genetic mapping of rpoD implicates the major sigma factor of Bacillus subtilis RNA polymerase in sporulation initiation. Mol. Gen. Genet. 201, 88–95.

    Article  PubMed  CAS  Google Scholar 

  9. Scott, N.W. and Dow, C.S. (1986) Purification and partial characterization of DNA-dependent RNA polymerase from Rhodomicrobium vannielii. J. Gen. Microbiol. 132, 1939–1949.

    PubMed  CAS  Google Scholar 

  10. Burgess, R.R. (1969) A new method for the large scale purification of Escherichia coli deoxyribonucleic acid-dependent ribonucleic acid polymerase. J. Biol. Chem. 244, 6160–6167.

    PubMed  CAS  Google Scholar 

  11. Burgess, R.R. and Jendrisak, J.J. (1975) A procedure for the rapid, large scale purification of Escherichia coli DNA-dependent RNA polymerase involving Polymin P precepitation and DNA cellulose chromatogaphy. Biochemistry 14, 4634–4638.

    Article  PubMed  CAS  Google Scholar 

  12. Alberts, B.M. and Herrick, G. (1971) DNA-cellulose chromatography Meth. Enzymol. 21, 198–217.

    Article  Google Scholar 

  13. Jaehning, J.A., Wiggs, J.L., and Chamberlin, M.J. (1979) Altered promoter selection by a novel form of Bacillus subtilis RNA polymerase. Proc. Natl. Acad. Sci. USA 76, 5470–5474.

    Article  PubMed  CAS  Google Scholar 

  14. Gonzalez, N., Wiggs, J., and Chamberlin, M.J. (1977) A single procedure for resolution of Escherichia coli RNA polymerase holoenzyme from core polymerase. Arch. Biochem. Biophys. 182, 404–408.

    Article  PubMed  CAS  Google Scholar 

  15. Gross, C., Engbaek, R., Flammang, T., and Burgess, R.R. (1976) Rapid micromethod for the purification of Escherichia coli ribonucleic acid polymerase and the preparation of bacterial extracts active in ribonucleic acid synthesis. J. Bacterial. 128, 382–389.

    CAS  Google Scholar 

  16. Haldenwang, W.G., Lang, N., and Losick, R. (1981) A sporulation-induced sigma-like regulatory protein from B. subtilis. Cell 23, 615–624.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. School of Life Sciences, Leicester Polytechnic, Leicester, UK

    N. W. Scott

Authors
  1. N. W. Scott
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. The Hatfield Polytechnic, Hatfield, Hertfordshire, UK

    John M. Walker

Rights and permissions

Reprints and permissions

Copyright information

© 1988 The Humana Press Inc.

About this protocol

Cite this protocol

Scott, N.W. (1988). Purification of DNA-Dependent RNA Polymerase from Eubacteria. In: Walker, J.M. (eds) New Protein Techniques. Methods in Molecular Biology™, vol 3. Humana Press. https://doi.org/10.1385/0-89603-126-8:135

Download citation

  • DOI: https://doi.org/10.1385/0-89603-126-8:135

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-126-5

  • Online ISBN: 978-1-59259-490-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation