Monitoring mitochondrial localization of dual localized proteins using a Bi-Genomic Mitochondrial-Split-GFP.
Dual-localized
Microscopy
Mitochondria
Protein import
Saccharomyces cerevisiae
Split-GFP
Journal
Methods in enzymology
ISSN: 1557-7988
Titre abrégé: Methods Enzymol
Pays: United States
ID NLM: 0212271
Informations de publication
Date de publication:
2024
2024
Historique:
medline:
26
10
2024
pubmed:
26
10
2024
entrez:
25
10
2024
Statut:
ppublish
Résumé
Even if a myriad of approaches has been developed to identify the subcellular localization of a protein, the easiest and fastest way remains to fuse the protein to Green Fluorescent Protein (GFP) and visualize its location using fluorescence microscopy. However, this strategy is not well suited to visualize the organellar pools of proteins that are simultaneously localized both in the cytosol and in organelles because the GFP signal of a cytosolic pool of the protein (cytosolic echoform) will inevitably mask or overlay the GFP signal of the organellar pool of the protein (organellar echoform). To solve this issue, we engineered a dedicated yeast strain expressing a Bi-Genomic Mitochondrial-Split-GFP. This split-GFP is bi-genomic because the first ten ß-strands of GFP (GFP
Identifiants
pubmed: 39455235
pii: S0076-6879(24)00364-1
doi: 10.1016/bs.mie.2024.07.028
pii:
doi:
Substances chimiques
Green Fluorescent Proteins
147336-22-9
Mitochondrial Proteins
0
Saccharomyces cerevisiae Proteins
0
Recombinant Fusion Proteins
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
75-95Informations de copyright
Copyright © 2024. Published by Elsevier Inc.