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An Efficient Gene Deletion Procedure for the Mushroom-Forming Basidiomycete Schizophyllum commune

Summary

This research developed an improved method for deleting genes in the mushroom-forming fungus Schizophyllum commune. The new technique makes it easier and more efficient to study gene functions in this important model organism. Impact on everyday life: • Helps scientists better understand how mushrooms grow and develop • Contributes to improving mushroom cultivation for food production • Advances our knowledge of fungal biology which is important for medicine and biotechnology • Could lead to improved production of useful compounds from fungi • May help develop better methods for controlling harmful fungi

Background

Gene deletion in Schizophyllum commune has historically been challenging due to low rates of homologous integration, requiring extensive screening to identify transformants with desired genotypes. Previous gene deletion attempts in S. commune showed varying success rates, with most genes being deleted at only about 3% efficiency.

Objective

To develop and validate an improved method for gene deletion in Schizophyllum commune using a newly constructed vector pDelcas that allows for directional cloning of flanking sequences and efficient screening of transformants.

Results

The new procedure achieved approximately 20-25% of nourseothricin resistant colonies showing phleomycin sensitivity, with about 4% of these having successful gene deletions. The system was validated on four different genes. Reducing phleomycin concentration to 5 μg/ml during screening eliminated up to 60% of false positives.

Conclusion

The developed pDelcas vector system combined with the improved PCR screening protocol provides an efficient method for gene deletion in S. commune. The procedure may also be applicable to other basidiomycete fungi, as S. commune expression cassettes have shown functionality in other species like Pycnoporus cinnabarinus, Agaricus bisporus, and Pleurotus ostreatus.
  • Published in:World Journal of Microbiology and Biotechnology,
  • Study Type:Laboratory Research,
  • Source: 10.1007/s11274-010-0356-0
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