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Related Experiment Videos

Protein expression using ubiquitin fusion and cleavage

R T Baker1

  • 1Molecular Genetics Group, John Curtin School of Medical Research, Australian National University, PO Box 334, Canberra, ACT 2601, Australia. Rohan.Baker@anu.edu.au

Current Opinion in Biotechnology
|October 1, 1996
PubMed
Summary

Protein expression is boosted by fusing proteins to ubiquitin, allowing controlled amino-terminal residues after cleavage. This technique is now more effective in bacterial and eukaryotic systems due to new ubiquitin-cleaving enzymes.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Protein Expression Systems

Background:

  • Fusion proteins offer enhanced protein yield and controlled N-terminal modifications.
  • Ubiquitin fusion technology facilitates the production of specific polypeptides.
  • Cleavage of ubiquitin from fusion proteins yields desired amino-terminal residues.

Purpose of the Study:

  • To highlight the advantages of using ubiquitin fusion for protein expression.
  • To discuss the impact of newly available ubiquitin-cleaving enzymes on this technique.
  • To demonstrate the applicability of ubiquitin fusion in diverse host systems.

Main Methods:

  • Utilizing ubiquitin as an N-terminal fusion partner for target proteins.
  • Employing cloned ubiquitin-cleaving enzymes for controlled cleavage.

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  • Implementing the technique in both bacterial and eukaryotic host systems.

    • Main Results:

    • Achieved a dramatic increase in the yield of expressed proteins and polypeptides.
    • Enabled precise control over the amino-terminal residue of the target protein post-cleavage.
    • Demonstrated successful application across different host systems.

    Conclusions:

    • Ubiquitin fusion is a powerful strategy for enhancing protein yield and producing proteins with specific N-termini.
    • The availability of cloned ubiquitin-cleaving enzymes significantly improves the efficiency and versatility of this method.
    • This approach is broadly applicable in both bacterial and eukaryotic expression systems for recombinant protein production.