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Science 15 January 1988:
Vol. 239. no. 4837, pp. 280 - 282
DOI: 10.1126/science.2827308
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Articles

Science, Vol 239, Issue 4837, 280-282
Copyright © 1988 by American Association for the Advancement of Science

articles

A general method for the chromosomal amplification of genes in yeast

JD Boeke, H Xu, and GR Fink

Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, MD 21205.

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.


THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
Identification and characterization of critical cis-acting sequences within the yeast Ty1 retrotransposon.
E. C. BOLTON, C. COOMBES, Y. EBY, M. CARDELL, and J. D. BOEKE (2005)
RNA 11, 308-322
   Abstract »    Full Text »    PDF »
Increase in Ty1 cDNA Recombination in Yeast sir4 Mutant Strains at High Temperature.
S. J. Radford, M. L. Boyle, C. J. Sheely, J. Graham, D. P. Haeusser, L. Zimmerman, and J. B. Keeney (2004)
Genetics 168, 89-101
   Abstract »    Full Text »    PDF »
Post-transcriptional Cosuppression of Ty1 Retrotransposition.
D. J. Garfinkel, K. Nyswaner, J. Wang, and J.-Y. Cho (2003)
Genetics 165, 83-99
   Abstract »    Full Text »    PDF »
An in vivo dual-luciferase assay system for studying translational recoding in the yeast Saccharomyces cerevisiae.
J. W. HARGER and J. D. DINMAN (2003)
RNA 9, 1019-1024
   Abstract »    Full Text »    PDF »
Transcriptional Interactions Between Yeast tRNA Genes, Flanking Genes and Ty Elements: A Genomic Point of View.
E. C. Bolton and J. D. Boeke (2003)
Genome Res. 13, 254-263
   Abstract »    Full Text »    PDF »
Ty1 Defect in Proteolysis at High Temperature.
J. F. Lawler Jr., D. P. Haeusser, A. Dull, J. D. Boeke, and J. B. Keeney (2002)
J. Virol. 76, 4233-4240
   Abstract »    Full Text »    PDF »
Frameshift Signal Transplantation and the Unambiguous Analysis of Mutations in the Yeast Retrotransposon Ty1 Gag-Pol Overlap Region.
J. F. Lawler Jr., G. V. Merkulov, and J. D. Boeke (2001)
J. Virol. 75, 6769-6775
   Abstract »    Full Text »    PDF »
A Ty1 Reverse Transcriptase Active-Site Aspartate Mutation Blocks Transposition but Not Polymerization.
O. Uzun and A. Gabriel (2001)
J. Virol. 75, 6337-6347
   Abstract »    Full Text »    PDF »
Isolation and Characterization of WHI3, a Size-Control Gene of Saccharomyces cerevisiae.
R. S. Nash, T. Volpe, and B. Futcher (2001)
Genetics 157, 1469-1480
   Abstract »    Full Text »
Ty1 Proteolytic Cleavage Sites Are Required for Transposition: All Sites Are Not Created Equal.
G. V. Merkulov, J. F. Lawler Jr., Y. Eby, and J. D. Boeke (2001)
J. Virol. 75, 638-644
   Abstract »    Full Text »    PDF »
Autocloning and Amplification of LIP2 in Yarrowia lipolytica.
G. Pignede, H.-J. Wang, F. Fudalej, M. Seman, C. Gaillardin, and J.-M. Nicaud (2000)
Appl. Envir. Microbiol. 66, 3283-3289
   Abstract »    Full Text »    PDF »
Invading the Yeast Nucleus: a Nuclear Localization Signal at the C Terminus of Ty1 Integrase Is Required for Transposition In Vivo.
M. A. Kenna, C. B. Brachmann, S. E. Devine, and J. D. Boeke (1998)
Mol. Cell. Biol. 18, 1115-1124
   Abstract »    Full Text »    PDF »
An unusual form of transcriptional silencing in yeast ribosomal DNA..
J S Smith and J D Boeke (1997)
Genes & Dev. 11, 241-254
   Abstract »    PDF »
Integration of the yeast retrotransposon Ty1 is targeted to regions upstream of genes transcribed by RNA polymerase III..
S E Devine and J D Boeke (1996)
Genes & Dev. 10, 620-633
   Abstract »    PDF »
A specific terminal structure is required for Ty1 transposition..
D J Eichinger and J D Boeke (1990)
Genes & Dev. 4, 324-330
   Abstract »    PDF »


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