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Fig. 2 | BMC Biology

Fig. 2

From: A tissue-specific protein purification approach in Caenorhabditis elegans identifies novel interaction partners of DLG-1/Discs large

Fig. 2

Schematic of the workflow. a The GTA tag is added to a gene of interest using recombineering. b Transgenic C. elegans strains expressing the GTA-tagged protein are generated by injection followed by gamma irradiation-mediated integration of the extrachromosomal array. c Transgenic strains are crossed with strains expressing BirA from a tissue-specific promoter, and with a genetic null mutant if appropriate. d The transgenic strains are grown in triplicate in liquid culture. e Affinity purification is performed on whole-animal lysates. The bait protein with any interacting proteins is subsequently cleaved off the beads by Tobacco etch virus (TEV) protease. f The samples are analyzed by tandem mass spectrometry (MS/MS) to identify the proteins they contain

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