A general method for the chromosomal amplification of genes in yeast.
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 GALI-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 TRPl 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.
Boeke
J D
JD
Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, MD 21205.
Xu
H
H
Fink
G R
GR
D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics; D004274:DNA, Recombinant; D005800:Genes, Fungal; D019868:Kanamycin Kinase; D021141:Nucleic Acid Amplification Techniques; D009693:Nucleic Acid Hybridization; D010770:Phosphotransferases / Q000235:genetics; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D014158:Transcription, Genetic; D014170:Transformation, Genetic
D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D004271:DNA, Fungal; D004274:DNA, Recombinant; D010770:Phosphotransferases; D019868:Kanamycin Kinase