Abstract
Many commonly used protein epitope tags are optimized for one or a few biological applications. The recently developed ALFA tag performs well across a broad range of cell biological applications including localization, biochemical, and functional studies but has not been available in budding yeast. This study engineers a modular plasmid-based platform to introduce ALFA tag on proteins of interest in budding yeast for detection via fluorescently tagged ALFA nanobodies. The toolkit offers excellent performance for quantitative live imaging, spatiotemporal analyses of proteins, protein kinetics, and immunoprecipitation studies, all with the same tag. These materials will be of broad interest to yeast biologists for simplified imaging and biochemical analyses. This preprint has been assigned the following badges: New Methods, New Materials. Read the preprint on bioRxiv ( Akhuli et al., 2022 ): https://doi.org/10.1101/2022.07.18.500560 .
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