Abstract
Labeling a protein of interest is widely used to examine its quantity, modification, localization, and dynamics in the budding yeast Saccharomyces cerevisiae. Fluorescent proteins and epitope tags are often used as protein fusion tags to study target proteins. One prevailing technique is to fuse these tags to a target gene at the precise chromosomal location via homologous recombination. Here we describe a protein labeling strategy based on the URA3 pop-in/pop-out and counterselection system to fuse a fluorescent protein or epitope tag scarlessly to a target protein at its native locus in S. cerevisiae.
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Acknowledgments
The authors wish to thank Huiping Guo for technical assistance and Huijun Xue for the development of method. This work was supported by the Natural Science Foundation of China operating grant 31270823 and Natural Sciences and Engineering Research Council of Canada Discovery Grant RGPIN-2019-05604 to WX.
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Wang, Q., Fu, Y.V., Xiao, W. (2021). Scarless Genomic Protein Labeling in Saccharomyces cerevisiae. In: Xiao, W. (eds) Yeast Protocols. Methods in Molecular Biology, vol 2196. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0868-5_6
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DOI: https://doi.org/10.1007/978-1-0716-0868-5_6
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