Assessment of GFP Tag Position on Protein Localization and Growth Fitness in Yeast
| Autor: | Zohar Avihou, Uri Weill, Ron Milo, Gat Krieger, Maya Schuldiner, Dan Davidi |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
0303 health sciences
Recombinant Fusion Proteins C-terminus Green Fluorescent Proteins Saccharomyces cerevisiae Computational biology Protein tag Biology Subcellular localization Protein subcellular localization prediction Article Yeast Green fluorescent protein Fungal Proteins Luminescent Proteins Protein Transport 03 medical and health sciences 0302 clinical medicine Structural Biology Genomic library Molecular Biology 030217 neurology & neurosurgery Function (biology) 030304 developmental biology |
| Zdroj: | J Mol Biol Journal of Molecular Biology |
| ISSN: | 0022-2836 |
| DOI: | 10.1016/j.jmb.2018.12.004 |
| Popis: | While protein tags are ubiquitously utilized in molecular biology, they harbor the potential to interfere with functional traits of their fusion counterparts. Systematic evaluation of the effect of protein tags on localization and function would promote accurate use of tags in experimental setups. Here we examine the effect of Green Fluorescent Protein (GFP) tagging at either the N or C terminus of budding yeast proteins on localization and functionality. We use a competition-based approach to decipher the relative fitness of two strains tagged on the same protein but on opposite termini and from that infer the correct, physiological localization for each protein and the optimal position for tagging. Our study provides a first of a kind systematic assessment of the effect of tags on the functionality of proteins and provides a step towards broad investigation of protein fusion libraries. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect