Mammalian expression vectors for metabolic biotinylation tandem affinity tagging by co-expression in cis of a mammalian codon-optimized BirA biotin ligase

Marina Ioannou, Dimitris N Papageorgiou, Vasily Ogryzko, John Strouboulis

    Research output: Contribution to journalArticlepeer-review

    3 Citations (Scopus)

    Abstract

    ΟBJECTIVE: To construct mammalian expression vectors for the N- or C-terminal tagging of proteins with a tandem affinity tag comprised of the biotinylatable Avi tag and of a triple FLAG tag.

    RESULTS: We constructed and tested by transient transfections mammalian expression vectors for the co-expression from a single plasmid of N- or C-terminally tagged proteins bearing a tandem affinity tag comprised of the biotinylatable Avi tag and of a triple FLAG tag separated by a tobacco etch virus (TEV) protease cleavage site, together with a mammalian codon-optimized BirA biotin ligase fused to green fluorescent protein. We also describe platform vectors for the N- or C-terminal AVI-TEV-FLAG tagging of any complementary DNA of choice. These vectors offer versatility and efficiency in the application of metabolic biotinylation tandem affinity tagging of nuclear proteins in mammalian cells.

    Original languageEnglish
    Pages (from-to)390
    JournalBMC Research Notes
    Volume11
    Issue number1
    DOIs
    Publication statusPublished - 14 Jun 2018

    Keywords

    • Affinity Labels
    • Animals
    • Biotinylation/methods
    • Genetic Vectors
    • HEK293 Cells
    • Humans
    • Mice
    • Plasmids
    • Rabbits
    • Rats

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