General purpose tagging vectors for fission yeast

Area of Science:

• Molecular Biology
• Yeast Genetics
• Protein Biochemistry

Background:

• Protein tagging is crucial for studying protein function and interactions.
• Existing methods for protein tagging in fission yeast may lack flexibility or efficiency.
• Schizosaccharomyces pombe is a key model organism for eukaryotic cell biology research.

Purpose of the Study:

• To create a versatile set of vectors for tagging proteins in Schizosaccharomyces pombe.
• To enable fusion of proteins with either a triple HA epitope or a GST domain.
• To provide options for N-terminal or C-terminal tagging and controllable gene expression.

Main Methods:

• Design and construction of a series of expression vectors for fission yeast.
• Inclusion of triple HA epitope and GST tags with modular design.
• Utilizing different versions of the nmt1 promoter for regulated gene expression.
• Demonstration of vector functionality using the Cdc19p protein.

Main Results:

• A novel family of vectors for Schizosaccharomyces pombe protein tagging was successfully developed.
• Vectors allow flexible N-terminal or C-terminal fusion with HA or GST tags.
• The nmt1 promoter enables tunable expression levels for tagged proteins.
• Functional tagged Cdc19p protein was expressed in fission yeast using the designed vectors.

Conclusions:

• The developed vector system provides a powerful and flexible tool for protein research in fission yeast.
• These vectors facilitate the study of protein localization, interactions, and function.
• The modular design simplifies the exchange of proteins between plasmids for comparative studies.