An improved tetracycline-inducible expression system for fission yeast

Area of Science:

• Molecular Biology
• Genetics
• Microbiology

Background:

• Gene expression manipulation is crucial for understanding gene function.
• The nmt1 promoter system in Schizosaccharomyces pombe has limitations like lag phase and unsuitability for non-dividing cells.

Purpose of the Study:

• To develop a novel tetracycline-inducible gene expression system for fission yeast.
• To overcome the limitations of the existing nmt1 promoter system.

Main Methods:

• Introduction of the enotetS promoter and its variants.
• Development of an integration plasmid containing the Tet-repressor cassette.
• Application in both dividing and non-dividing fission yeast cells.

Main Results:

• The enotetS promoter shows a higher induction ratio and no lag compared to nmt1.
• Four weakened enotetS variants provide a tunable expression range.
• The system supports gene expression control in non-dividing cells, enabling synchronous meiosis induction.
• Conditional gene silencing and loss-of-function mutant generation are possible.

Conclusions:

• The new tetracycline-inducible system offers a versatile and powerful tool for gene expression studies in Schizosaccharomyces pombe.
• It enhances experimental possibilities, including conditional gene silencing and synchronous meiosis induction.