Skip to main content

Protein Extraction, Fractionation, and Purification From Cyanobacteria

  • Protocol
Circadian Rhythms

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 362))

Abstract

This chapter deals with methods of protein extraction from cyanobacterial cells based on work in the circadian model organism Synechococcus elongatus PCC 7942. Some of these techniques have already been used successfully for analysis of circadian rhythms in cyanobacteria, whereas others are heretofore unpublished, but may yield exciting results in the near future.

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

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 169.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 219.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 219.99
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

Institutional subscriptions

References

  1. Nakahira, Y., Katayama, M., Miyashita, H., et al. (2004) Global gene repression by KaiC as a master process of prokaryotic circadian system. Proc. Natl. Acad. Sci. USA 101, 881–885.

    Article  CAS  PubMed  Google Scholar 

  2. Ouyang, Y., Andersson, C. R., Kondo, T., Golden, S. S., and Johnson, C. H. (1998) Resonating circadian clocks enhance fitness in cyanobacteria. Proc. Natl. Acad. Sci. USA 95, 8660–8664.

    Article  CAS  PubMed  Google Scholar 

  3. Kageyama, H., Kondo, T., and Iwasaki, H. (2003) Circadian formation of clock protein complexes by KaiA, KaiB, KaiC, and SasA in cyanobacteria. J. Biol. Chem. 278, 2388–2395.

    Article  CAS  PubMed  Google Scholar 

  4. Iwasaki, H., Taniguchi, Y., Ishiura, M., and Kondo, T. (1999) Physical interactions among circadian clock proteins KaiA, KaiB and KaiC in cyanobacteria. EMBO J. 18, 1137–1145.

    Article  CAS  PubMed  Google Scholar 

  5. Iwasaki, H., Nishiwaki, T., Kitayama, Y., Nakajima, M., and Kondo, T. (2002) KaiA-stimulated KaiC phosphorylation in circadian timing loops in cyanobacteria. Proc. Natl. Acad. Sci. USA 99, 15,788–15,793.

    Article  CAS  PubMed  Google Scholar 

  6. Nair, U., Ditty, J. L., Min, H., and Golden, S. S. (2002) Roles for sigma factors in global circadian regulation of the cyanobacterial genome. J. Bacteriol. 184, 3530–3538.

    Article  CAS  PubMed  Google Scholar 

  7. Mutsuda, M., Michel, K.-P., Zhang, X., Montgomery, B. L., and Golden, S. S. (2003) Biochemical properties of CikA, an unusual phytochrome-like histidine protein kinase that resets the circadian clock in Synechococcus elongatus PCC 7942. J. Biol. Chem. 278, 19,102–19,110.

    Article  CAS  PubMed  Google Scholar 

  8. Kitayama, Y., Iwasaki, H., Nishiwaki, T., and Kondo, T. (2003) KaiB functions as an attenuator of KaiC phosphorylation in the cyanobacteria circadian clock system. EMBO J. 22, 1–8.

    Article  Google Scholar 

  9. Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual. 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.

    Google Scholar 

  10. The QIAexpressionist: A Handbook for High-Level Expression and Purification of 6x His-Tagged Proteins. 5th ed. Qiagen, Valencia, CA.

    Google Scholar 

  11. Lowry, O. H., Rosebrough, N. J., Farr, A. L., and Randall, R. J. (1951) Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193, 265–275.

    CAS  PubMed  Google Scholar 

  12. Fulda, S., Mikkat, S., Schroder, W., and Hagemann, M. (1999) Isolation of saltinduced periplasmic proteins from Synechocystis sp. strain PCC 6803. Arch. Microbiol. 171, 214–217.

    Article  CAS  PubMed  Google Scholar 

  13. Norling, B., Zak, E., Andersson, B., and Pakrasi, H. (1998) 2D-isolation of pure plasma and thylakoid membranes from the cyanobacterium Synechocystis sp. PCC 6803. FEBS Lett. 436, 189–192.

    Article  CAS  PubMed  Google Scholar 

  14. Zak, E., Norling, B., Andersson, B., and Pakrasi, H. B. (1999) Subcellular localization of the BtpA protein in the cyanobacterium Synechocystis sp. PCC 6803. Eur. J. Biochem. 261, 311–316.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2007 Humana Press Inc.

About this protocol

Cite this protocol

Ivleva, N.B., Golden, S.S. (2007). Protein Extraction, Fractionation, and Purification From Cyanobacteria. In: Rosato, E. (eds) Circadian Rhythms. Methods in Molecular Biology™, vol 362. Humana Press. https://doi.org/10.1007/978-1-59745-257-1_26

Download citation

  • DOI: https://doi.org/10.1007/978-1-59745-257-1_26

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-417-3

  • Online ISBN: 978-1-59745-257-1

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics