| Autor: |
Fizza Askari, Rupinder Kaur |
| Jazyk: |
angličtina |
| Rok vydání: |
2024 |
| Předmět: |
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| Zdroj: |
STAR Protocols, Vol 5, Iss 1, Pp 102759- (2024) |
| Druh dokumentu: |
article |
| ISSN: |
2666-1667 |
| DOI: |
10.1016/j.xpro.2023.102759 |
| Popis: |
Summary: Phosphatidylinositol 3-phosphate (PI3P) levels govern membrane trafficking in Candida glabrata. Here, we present a confocal imaging-based protocol for PI3P localization analysis using the GFP-FYVE (found in Fab1, YOTB, Vac1, and EEA1) fusion protein. We describe steps for cloning the FYVE domain into the GFP-containing vector backbone, transforming FYVE-GFP into C. glabrata, and preparing slides with FYVE-GFP-expressing C. glabrata cells. We then detail procedures for acquiring and analyzing images and quantifying signal data. This protocol is adaptable to subcellular localization analysis of other low-abundant lipid and protein molecules.For complete details on the use and execution of this protocol, please refer to Askari et al. (2023).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| Databáze: |
Directory of Open Access Journals |
| Externí odkaz: |
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