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Genetics

Title:		Ty3 transposes in mating populations of yeast: a novel transposition assay for Ty3
Author(s):		Kinsey PT; Sandmeyer SB;
Address:		"Department of Microbiology and Molecular Genetics, College of Medicine, University of California, Irvine 92717"
Journal Title:		Genetics
Year:		1995
Volume:		139
Issue:		1
Page Number:		81 - 94
DOI:		10.1093/genetics/139.1.81
ISSN/ISBN:		0016-6731 (Print) 0016-6731 (Linking)
Abstract:		"Ty3 is a retrotransposon of Saccharomyces cerevisiae that integrates just upstream of the transcription initiation site of genes transcribed by RNA polymerase III. Ty3 transcription is pheromone-inducible in haploid cells and is mating-type regulated in diploid cells. The specificity of Ty3 integration was exploited in the design of a novel target into which transposition of Ty3 elements could be selected. The target plasmid contains divergently oriented tRNA genes with 19 base pairs separating the two tRNA gene coding sequences. An inactive ochre suppressor tRNA(Tyr) gene with a modified transcription initiation region was used as the selectable marker and a tRNA(Val) (AAC) gene was used to direct Ty3 integration into the transcription initiation region of the suppressor tRNA(Tyr) gene. Integration of Ty3 activated expression of the suppressor tRNA gene, which resulted in suppression of ochre nonsense alleles ade2-101(0) and lys2-1(0) and allowed cell growth on selective medium. Based on the activity of this target, Ty3, under control of a galactose-inducible promoter and present on a high copy-number plasmid, was estimated to transpose into the genome at a rate of 5.6 x 10(-3) per cell division. We show here that induction of Ty3 transcription from its natural promoter results in transposition. Ty3 elements in strains of the a or alpha mating-type transposed efficiently to target plasmids in cells of the opposite mating-type. Thus, natural transposition of Ty3 is regulated temporally to occur in mating populations"
Keywords:		"Adenine/metabolism Base Sequence Genetic Markers Lysine/metabolism Molecular Sequence Data Plasmids/genetics RNA, Transfer, Tyr/genetics RNA, Transfer, Val/genetics Reproduction Retroelements/*genetics Saccharomyces cerevisiae/*genetics Selection, Genetic;"
Notes:		"MedlineKinsey, P T Sandmeyer, S B eng GM-0713417/GM/NIGMS NIH HHS/ GM-33281/GM/NIGMS NIH HHS/ Research Support, U.S. Gov't, P.H.S. 1995/01/01 Genetics. 1995 Jan; 139(1):81-94. doi: 10.1093/genetics/139.1.81"