Fig. 1. TurboID is an efficient biotin ligase in Xenopus embryos. (A) Diagrams of HA-tagged TurboID and BioID2. (B) Assay for biotin ligase mediated-biotinylation. Two-cell Xenopus embryos were injected with mRNA encoding either HA-tagged BioID2 or HA-tagged TurboID with biotin supplemented in the culture media at indicated concentrations. When embryos reached stage 7, protein was extracted for analysis of biotinylation. (C) Immunoblot analysis with an anti-HA antibody was used to monitor the expression of injected mRNA (Lanes 1â3, top panel). Streptavidin-HRP assayed self-biotinylation of each enzyme as a readout of enzyme activity (Lanes 1â3, middle panel). Blots were stripped and re-probed with actin to assess sample loading (Lanes 1â3, bottom panel). Open triangles indicate where biotinylation of BioID2 is to be expected. Figure is representative of three biological replicates.
Image published in: Kanzler CR et al. (2022)
Copyright © 2022. Image reproduced with permission of the Publisher, Elsevier B. V.
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