One-step purification of E. coli elongation factor Tu.

Charlotte Rohde Knudsen, Brian F. C. Clark, B Degn, Ove Wiborg

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    Abstract

    The tuf A gene, encoding the E. coli elongation factor Tu, was cloned in the pGEX gene fusion system. Upon expression EF-Tu is fused to glutathione-S-transferase serving as a purification handle with affinity for glutathione immobilised on agarose. This allows purification of EF-Tu in a one-step procedure. The construct was designed in order to make possible the release of authentic EF-Tu by cleaving the fusion protein with the protease factor Xa.
    Udgivelsesdato: 1992-Oct
    Original languageEnglish
    JournalBiochemistry international
    Volume28
    Issue2
    Pages (from-to)353-62
    Number of pages9
    ISSN0158-5231
    Publication statusPublished - 1993

    Keywords

    • Amino Acid Sequence
    • Anti-Bacterial Agents
    • Cloning, Molecular
    • Electrophoresis, Polyacrylamide Gel
    • Escherichia coli
    • Factor Xa
    • Gene Expression
    • Glutathione Transferase
    • Molecular Sequence Data
    • Mutagenesis, Site-Directed
    • Peptide Elongation Factor Tu
    • Plasmids
    • Pyridones
    • Recombinant Fusion Proteins

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