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Updated: Feb 9, 2026

Expression and Purification of Mammalian Bestrophin Ion Channels
Published on: August 2, 2018
Mammalian expression vectors for metabolic biotinylation tandem affinity tagging by co-expression in cis of a
Marina Ioannou1, Dimitris N Papageorgiou2,3, Vasily Ogryzko4
1Institute of Molecular Biology and Biotechnology, Foundation of Research & Technology Hellas, 100 Nikolaou Plastira Street, 70013, Heraklion, Crete, Greece.
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.
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