Analysis of protein function in clinical C. albicans isolates

Area of Science:

• Microbiology
• Molecular Biology
• Genetics

Background:

• Clinical isolates of Candida albicans are prototrophic, limiting genetic manipulation via nutritional markers.
• Existing genetic tools are insufficient for comprehensive gene function studies in diverse C. albicans strains.

Purpose of the Study:

• To develop a versatile set of plasmids for genetic manipulation in both clinical and laboratory strains of C. albicans.
• To facilitate PCR-mediated construction of epitope-tagged proteins and conditional gene expression.
• To enable in vivo studies of protein co-expression and co-localization.

Main Methods:

• Construction of novel plasmids utilizing the NAT1 drug resistance marker.
• Development of plasmids for C-terminal epitope tagging (HA-NAT1, MYC-NAT1).
• Creation of plasmids for conditional protein expression (NAT1-pMet3-GFP) and internal GFP tagging (GF-NAT1-FP).
• Construction of plasmids for dual-labeling studies (GFP-NAT1, RFP-NAT1, M-Cherry-NAT1).

Main Results:

• Successfully constructed and validated a new set of plasmids carrying the NAT1 resistance marker.
• Demonstrated utility for C-terminal epitope tagging, conditional expression, and internal protein tagging.
• Developed dual-labeling vectors for studying protein co-expression and co-localization.
• These vectors are effective in both clinical and laboratory C. albicans strains.

Conclusions:

• The developed plasmids represent a valuable toolkit for advancing the study of gene function in C. albicans.
• These genetic tools enhance the ability to investigate diverse aspects of gene expression and protein interactions.
• The NAT1 marker-based plasmids offer broad applicability across different C. albicans strain types.