Fig. 7
Retrotransposition of the TRE5-A population in agnC overexpressing strains. a Semi-quantitative RT-PCR analysis of RNA from DH1 and three independent DH1[agnC GA] mutants. Note that agnC was overexpressed comapred to DH1. Expression of the neighboring gene DDB_G0271884 (compare Figure 5) was not affected, whereas expression of TRE5-A ORF1 was reduced in the agnC GA mutants. Expression of the house-keeping gene gpdA is shown as control. NTC: no template control. b DH1 and DH1[agnC GA] cells were transformed with either the empty TRE trap gene (i.e., no tRNA gene inserted in the intron) or the TRE trap gene containing a Val UAC tRNA gene. In this TRE trap assay, one clone of DH1 transformants and two independent clones of DH1[agnC GA] cells (clones #1 and #2) were analyzed for TRE5-A retrotransposition. For each strain, five plates containing 107 cells each were prepared. Cells were cultured in FM medium containing 5-FOA and uracil until clones appeared. Clones were counted and results are presented as retrotransposition frequency ± SD. ***p < 0.001 (Student’s t-test). This assay was reproduced once with similar results