|
English
|
正體中文
|
简体中文
|
Items with full text/Total items : 17934/22951 (78%)
Visitors : 7404507
Online Users : 107
|
|
|
Loading...
|
Please use this identifier to cite or link to this item:
https://ir.csmu.edu.tw:8080/ir/handle/310902500/20635
|
Title: | Establishment of tetracycline-regulated bimolecular fluorescence complementation assay to detect protein-protein interactions in Candida albicans |
Authors: | Lai, W.-C. Sun, H.S. Shieh, J.-C. |
Keywords: | Candida albicans;Virulence;Human fungal |
Date: | 2020-12-01 |
Issue Date: | 2020-03-11T04:27:04Z (UTC)
|
Publisher: | Scientific Reports |
ISSN: | 2045-2322 |
Abstract: | To visualize protein-protein interactions in Candida albicans with the bimolecular fluorescence complementation (BiFC) approach, we created a Tet-on system with the plasmids pWTN1 and pWTN2. Both plasmids bear a hygromycin B-resistant marker (CaHygB) that is compatible with the original Tet-on plasmid pNIM1, which carries a nourseothricin-resistant marker (CaSAT1). By using GFPmut2 and mCherry as reporters, we found that the two complementary Tet-on plasmids act synergistically in C. albicans with doxycycline in a dose-dependent manner and that expression of the fusion proteins, CaCdc11-GFPmut2 and mCherry-CaCdc10, derived from this system, is septum targeted. Furthermore, to allow detection of protein-protein interactions with the reassembly of a split fluorescent protein, we incorporated mCherry into our system. We generated pWTN1-RN and pNIM1-RC, which express the N-terminus (amino acids 1–159) and C-terminus (amino acids 160–237) of mCherry, respectively. To verify BiFC with mCherry, we created the pWTN1-CDC42-RN (or pWTN1-RN-CDC42) and pNIM1-RC-RDI1 plasmids. C. albicans cells containing these plasmids treated with doxycycline co-expressed the N- and C-terminal fragments of mCherry either N-terminally or C-terminally fused with CaCdc42 and CaRdi1, respectively, and the CaCdc42-CaRdi1 interaction reconstituted a functional form of mCherry. The establishment of this Tet-on-based BiFC system in C. albicans should facilitate the exploration of protein-protein interactions under a variety of conditions. © 2020, The Author(s). |
URI: | https://ir.csmu.edu.tw:8080/ir/handle/310902500/20635 |
Relation: | Scientific Reports, Volume 10, Issue 1, no. 2936 |
Appears in Collections: | [生化微生物免疫研究所] 期刊論文
|
Files in This Item:
File |
Description |
Size | Format | |
Establishment-of-tetracyclineregulated-bimolecular-fluorescence-complementation-assay-to-detect-proteinprotein-interactions-in-Candida-albicans2020Scientific-Reports.pdf | | 2778Kb | Adobe PDF | 175 | View/Open |
|
All items in CSMUIR are protected by copyright, with all rights reserved.
|