Skip to main content
SpringerLink
Log in

In Vitro Translation Assays for Selenocysteine Insertion

  • Protocol
  • First Online:
Selenoproteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1661))

Abstract

The molecular characterization of the protein and RNA factors that are required for Sec incorporation in mammals has been largely carried out using in vitro translation systems specifically modified for this purpose. This chapter outlines the various systems and modifications that have been used to decipher the mechanism of Sec incorporation.

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

Access this chapter

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 139.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 179.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 179.99
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. Pelham HR, Jackson RJ (1976) An efficient mRNA-dependent translation system from reticulocyte lysates. Eur J Biochem 67:247–256

    Article  CAS  PubMed  Google Scholar 

  2. Anderson CW, Straus JW, Dudock BS (1983) Preparation of a cell-free protein-synthesizing system from wheat germ. Methods Enzymol 101:635–644

    Article  CAS  PubMed  Google Scholar 

  3. Lesoon A, Mehta A, Singh R, Chisolm GM, Driscoll DM (1997) An RNA-binding protein recognizes a mammalian selenocysteine insertion sequence element required for cotranslational incorporation of selenocysteine. Mol Cell Biol 17:1977–1985

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Copeland PR, Driscoll DM (1999) Purification, redox sensitivity, and RNA binding properties of SECIS-binding protein 2, a protein involved in selenoprotein biosynthesis. J Biol Chem 274:25447–25454

    Article  CAS  PubMed  Google Scholar 

  5. Copeland PR, Fletcher JE, Carlson BA, Hatfield DL, Driscoll DM (2000) A novel RNA binding protein, SBP2, is required for the translation of mammalian selenoprotein mRNAs. EMBO J 19:306–314

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Kinzy SA, Caban K, Copeland PR (2005) Characterization of the SECIS binding protein 2 complex required for the co-translational insertion of selenocysteine in mammals. Nucleic Acids Res 33:5172–5180

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Gupta N, Demong LW, Banda S, Copeland PR (2013) Reconstitution of selenocysteine incorporation reveals intrinsic regulation by SECIS elements. J Mol Biol 425:2415–2422

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

This work was supported by NIH grants GM077073 and HD083616 to PRC.

Author information

Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Rutgers-Robert Wood Johnson Medical School, 675 Hoes Lane, Piscataway, NJ, 08854, USA

    Mark H. Pinkerton & Paul R. Copeland

Authors
  1. Mark H. Pinkerton
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Paul R. Copeland
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Paul R. Copeland .

Editor information

Editors and Affiliations

  1. Centre International de Recherche en Infectiologie, CIRI, Lyon, France

    Laurent Chavatte

Rights and permissions

Reprints and permissions

Copyright information

© 2018 Springer Science+Business Media LLC

About this protocol

Cite this protocol

Pinkerton, M.H., Copeland, P.R. (2018). In Vitro Translation Assays for Selenocysteine Insertion. In: Chavatte, L. (eds) Selenoproteins. Methods in Molecular Biology, vol 1661. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7258-6_7

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-7258-6_7

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7257-9

  • Online ISBN: 978-1-4939-7258-6

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation