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Genetics, Vol 120, 95-108, Copyright © 1988
INVESTIGATIONS |
Transposon Tagging Using Ty Elements in Yeast
D. J. Garfinkel, M. F. Mastrangelo, N. J. Sanders, B. K. Shafer and J. N. Strathern
Bionetics Research, Inc., Basic Research Program, National Cancer Institute-Frederick Cancer Research Facility, Frederick, Maryland 21701
We have used the ability to induce high levels of Ty transposition to develop a method for transposon mutagenesis in Saccharomyces cerevisiae. To facilitate genetic and molecular analysis, we have constructed GAL1-promoted TyH3 or Ty917 elements that contain unique cloning sites, and marked these elements with selectable genes. These genes include the yeast HIS3 gene, and the plasmid PiAN7 containing the Tn903 NEO gene. The marked Ty elements retain their ability to transpose, to mutate the LYS2, LYS5, or STE2 genes, and to activate the promoterless his3{Delta}4 target gene. Ty elements containing selectable genes are also useful in strain construction, in chromosomal mapping, and in gene cloning strategies.
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