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A split ALFA tag-nanobody system for protein localization and proximity proteomics in mycobacteria.

Fay A, Kurland AP, Li Z, Monetti M, Johnson JR, Glickman MS
mBio. 2025 Aug 13;16(8):e0097125. doi: 10.1128/mbio.00971-25. Epub 2025 Jun 27. (Link opens in a new window) PubMed (Link opens in a new window) Article

Plasmids from Article

ID Plasmid Purpose
252903pAJF1120Tet-inducible GFP-NBALFA fusion plasmid. Contains oriM and kanamycin-resistance cassette.
252904pAJF11243' rpoC(MSMEG_1368)-ALFA for rpoC-ALFA in M. smegmatis via single crossover. Encodes hygromycin-resistance cassette.
252906pAJF1131T7-pelB-NBALFA-LE-6his amp (for purification of his-tagged NBALFA from E. coli)
252907pAJF1137Tet-inducible TurboID-NBALFA fusion plasmid. Contains oriM (mycobacterial origin of replication) and kanamycin-resistance cassette.
252908pAJF1139For constitutive expression of GFP-NBALFA. Contains oriM (mycobacterial origin of replication) and kanamycin-resistance cassette.
252909pAJF1141For constitutive expression of GFP-NBALFA. Contains oriM (mycobacterial origin of replication) and kanamycin-resistance cassette.

Antibodies from Article