Single-step selection for Ty1 element retrotransposition.

AUTOR(ES)

Curcio, M J

RESUMO

The yeast retrotransposon Ty1 has been tagged with a reporter gene that allows selection of RNA-mediated transposition events and is applicable to the study of retroelements in other organisms. The reporter gene is a yeast HIS3 gene interrupted by an artificial intron (AI) in the antisense orientation. The HIS3AI sequences were inserted into a Ty1 element such that the intron is on the sense strand of the Ty1 element; therefore, splicing and retrotransposition of marked Ty1 transcripts can give rise to His+ cells. Fusion of the Ty1-H3mHIS3AI element to the inducible GAL1 promoter resulted in a high frequency of histidine prototrophs upon galactose induction. Moreover, spontaneous His+ revertants derived from strains containing genomic TymHIS3AI elements are a result of retrotransposition. By using this assay, we estimated the Ty1 transposition rate to be between 3 x 10(-7) and 1 x 10(-5) transpositions per Ty1 element per generation. Variations in the transposition rate of individual Ty1 elements are correlated with the relative abundance of their transcripts.

Documentos Relacionados

• Replication infidelity during a single cycle of Ty1 retrotransposition.
• The primer tRNA sequence is not inherited during Ty1 retrotransposition.
• Relationship between RNA Lariat Debranching and Ty1 Element Retrotransposition
• A Rare Trna-Arg(ccu) That Regulates Ty1 Element Ribosomal Frameshifting Is Essential for Ty1 Retrotransposition in Saccharomyces Cerevisiae
• Single-step direct PCR amplification from solid tissues.