Assessment of GFP Tag Position on Protein Localization and Growth Fitness in Yeast.
Cellular fitness
Genomic libraries
Green fluorescent protein
Protein localization
Protein tag
Journal
Journal of molecular biology
ISSN: 1089-8638
Titre abrégé: J Mol Biol
Pays: Netherlands
ID NLM: 2985088R
Informations de publication
Date de publication:
01 02 2019
01 02 2019
Historique:
received:
20
06
2018
revised:
05
11
2018
accepted:
05
12
2018
pubmed:
15
12
2018
medline:
3
3
2020
entrez:
15
12
2018
Statut:
ppublish
Résumé
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 function would promote accurate use of tags in experimental setups. Here we examine the effect of green fluorescent protein tagging at either the N or C terminus of budding yeast proteins on subcellular 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 toward broad investigation of protein fusion libraries.
Identifiants
pubmed: 30550779
pii: S0022-2836(18)30648-X
doi: 10.1016/j.jmb.2018.12.004
pmc: PMC7611381
mid: EMS130947
pii:
doi:
Substances chimiques
Fungal Proteins
0
Luminescent Proteins
0
Recombinant Fusion Proteins
0
Green Fluorescent Proteins
147336-22-9
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
636-641Subventions
Organisme : European Research Council
ID : 646604
Pays : International
Informations de copyright
Copyright © 2018 The Authors. Published by Elsevier Ltd.. All rights reserved.
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