A general method for the chromosomal amplification of genes in yeast

Jef D. Boeke, Hua Xu, Gerald R. Fink

    Research output: Contribution to journalArticle

    Abstract

    The yeast retrotransposon Ty can be used to insert multiple copies of a gene at new sites in the genome. The gene of interest is inserted into a GAL1-Ty fusion construct; the entire "amplification cassette" is then introduced into yeast on a high copy number plasmid vector. Transposition of the Ty element carrying the gene occurs at multiple sites in the genome. Two genes, a bacterial neomycin phosphotransferase gene and the yeast TRP1 gene, were amplified in this way. Although the amplified genes were about 1 kilobase in length, they were amplified to about the same extent as a 40-base pair segment. The benefit of this "shotgun" approach is that amplification can be achieved in one set of manipulations.

    Original languageEnglish (US)
    Pages (from-to)280-282
    Number of pages3
    JournalScience
    Volume239
    Issue number4837
    DOIs
    StatePublished - 1988

    ASJC Scopus subject areas

    • General

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