Saccharomyces cerevisiae::Ty1-URA3 amplicon reads
Saccharomyces cerevisiae
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.
strain
BY4741
isolation_source
Laboratory
collection_date
2013-04-01
geo_loc_name
USA: Austin
sample_type
Cell culture
temp
30C
BioSampleModel
Microbe, viral or environmental