The SP-GFP-BirA* construct is located in the PV and is able to biotinylate proteins in this compartment. Immune fluorescence assay (IFA) of SP-GFP-BirA* expressing parasites grown in the presence of biotin show co-localisation of biotinylated protein (detected using streptavidin) with GFPBirA* (upper panel) and ETRAMP5 (lower panel). To test whether BirA* can biotinylate proteins in the PV we grew the SP-GFP-BirA* expressing parasites in the presence of biotin. Immuno and streptavidin-fluorescence assays showed that this resulted in biotinylation of proteins in the parasite periphery and that the detected biotin co-localized with the SP-GFP-BirA* construct and with the PVM marker ETRAMP5.Khosh-Naucke M, Becker J, Mesén-Ramírez P, Kiani P, Birnbaum J, Fröhlke U, Jonscher E, Schlüter H, Spielmann T. Identification of novel parasitophorous vacuole proteins in P. falciparum parasites using BioID. Int J Med Microbiol. 2017 Jul 27. [Epub ahead of print]