Kinetics of transcription and splicing of RiboSys reporter transcripts in tetOFF and tetON strains. A time course is shown of the levels of pre-mRNA (pre), lariat intron-exon2 splicing intermediate (lar-exon2; measured by 3′SS assay and subtracting the amount of unspliced pre-mRNA) (see Fig.1), exon2 (exon) and spliced mRNA in copies per cell. (A–C) tetOFF cultures were grown in SD medium to midlog phase (OD600 0.5) with transcription repressed by the presence of dox (4 μg/ mL). After 90 min of transcriptional repression, cells were harvested by centrifugation and resuspended into medium without dox (time 0). (D–F) tetON cultures (YIK91 with various Ribo1 reporters integrated at the his3 locus) were grown in SD minimal medium (-trp) with doxycline (4 μg/ mL) present to induce transcription. (A,D) Ribo1; (B,E) 5′SSRibo1; and (C,F) 3′SSRibo1. In each panel the data represent three experiments, each assayed in triplicate, and error bars indicate standard deviation.
