organism: Saccharomyces cerevisiae
description: In order to increase achievable library size during directed evolution, we aimed to harness the mutagenic capacity of Ty1 reverse transcriptase. Specifically, we inserted the URA3 gene into a galactose-activated Ty1 retrotransposon in the reverse orientation relative to Ty1 transcription, and transformed a plasmid containing this construct into S. cerevisiae BY4741. Then, by next-generation sequencing of a PCR-amplified locus of the URA3 gene after incubation with either glucose or galactose, we were able to measure conferred error rate by Ty1RT in this system.
attributes.tag: strain
attributes.value: BY4741
attributes.tag: isolation_source
attributes.value: Laboratory
attributes.tag: collection_date
attributes.value: 2013-04-01
attributes.tag: geo_loc_name
attributes.value: USA: Austin
attributes.tag: sample_type
attributes.value: Cell culture
attributes.tag: temp
attributes.value: 30C
attributes.tag: BioSampleModel
attributes.value: Microbe, viral or environmental