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Related Experiment Videos

Transformation of yeast.

A Hinnen, J B Hicks, G R Fink

    Proceedings of the National Academy of Sciences of the United States of America
    |April 1, 1978
    PubMed
    Summary
    This summary is machine-generated.

    Yeast strains were genetically modified using a ColE1 plasmid containing the LEU2 gene. The study demonstrates that both bacterial plasmid and yeast DNA sequences integrate into yeast chromosomes, affecting gene function and location.

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    Area of Science:

    • Molecular Biology
    • Yeast Genetics
    • Genetic Engineering

    Background:

    • Stable yeast strains with the leu2- mutation were used.
    • Transformation was achieved using a chimeric ColE1 plasmid carrying the yeast leu2 gene.

    Purpose of the Study:

    • To demonstrate the presence of transforming DNA in the yeast genome.
    • To determine the arrangement of integrated yeast and bacterial DNA sequences.
    • To analyze the genetic behavior and chromosomal locations of integrated sequences.

    Main Methods:

    • Hybridization techniques
    • Restriction endonuclease mapping
    • Genetic linkage analysis

    Main Results:

    • The transforming DNA, including ColE1 plasmid and yeast sequences, integrated into yeast chromosomes.

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  • Integration events were either additive or substitutive.
  • Integrated bacterial plasmid sequences followed Mendelian inheritance.
  • Transforming sequences integrated into the leu2 region and other chromosomal locations.

    • Conclusions:

    • Chimeric plasmids can facilitate the integration of bacterial and yeast DNA into the yeast genome.
    • Integrated DNA sequences can alter yeast genetic makeup and exhibit stable inheritance.
    • Integration is not restricted to the target gene locus, indicating potential for genomic alterations.