Additional file 1: Figure S1. Restriction maps of plasmid Avi-TEV-3xFLAG_pBS SK (A) and of plasmid 3xFLAG-TEV-Avi_pBS KS (B)

Dimitris Papageorgiou (5392694)

John Strouboulis (63634)

Marina Ioannou (5392697)

Vasily Ogryzko (16707)

BMC Research Notes

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

Marina Ioannou

Dimitris N. Papageorgiou

Vasily Ogryzko

John Strouboulis

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

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MOESM1 of Mammalian expression vectors for metabolic biotinylation tandem affinity tagging by co-expression in cis of a mammalian codon-optimized BirA biotin ligase

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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.</p

Ioannou, Marina

Papageorgiou, Dimitris N

Ogryzko, Vasily

Strouboulis, John

King's Research Portal

https://figshare.com/articles/MOESM1_of_Mammalian_expression_vectors_for_metabolic_biotinylation_tandem_affinity_tagging_by_co-expression_in_cis_of_a_mammalian_codon-optimized_BirA_biotin_ligase/6529622

MOESM1 of Mammalian expression vectors for metabolic biotinylation tandem affinity tagging by co-expression in cis of a mammalian codon-optimized BirA biotin ligase

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