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Related Experiment Videos

Transposon tagging using Ty elements in yeast.

D J Garfinkel1, M F Mastrangelo, N J Sanders

  • 1Bionetics Research, Inc., National Cancer Institute-Frederick Cancer Research Facility, Maryland 21701.

Genetics
|September 1, 1988
PubMed
Summary

Researchers developed a new transposon mutagenesis method in Saccharomyces cerevisiae using Ty elements. This technique facilitates genetic analysis by incorporating selectable genes into Ty elements for various applications.

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Area of Science:

  • Molecular Biology
  • Yeast Genetics
  • Transposon Mutagenesis

Background:

  • Ty elements are mobile genetic elements in Saccharomyces cerevisiae.
  • Transposon mutagenesis is a powerful tool for genetic analysis.
  • Previous methods for Ty transposition had limitations.

Purpose of the Study:

  • To develop an efficient method for transposon mutagenesis in Saccharomyces cerevisiae.
  • To create marked Ty elements for enhanced genetic and molecular analysis.
  • To facilitate strain construction, chromosomal mapping, and gene cloning.

Main Methods:

  • Utilized inducible high-level Ty transposition.
  • Constructed GAL1-promoted TyH3 and Ty917 elements with unique cloning sites.
  • Marked Ty elements with selectable genes (HIS3 and NEO).

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Main Results:

  • Marked Ty elements retained transposition ability.
  • Successfully mutated LYS2, LYS5, and STE2 genes.
  • Activated the promoterless his3 delta 4 target gene.
  • Demonstrated utility in strain construction, mapping, and cloning.

Conclusions:

  • The developed method provides a robust tool for transposon mutagenesis in yeast.
  • Marked Ty elements offer versatility for various genetic and molecular applications.
  • This approach enhances the study of Saccharomyces cerevisiae genetics and genome organization.