Figure 1.
The P5SM splicing cassette functions in the eGFP coding sequence. (A) Schematic representation of the P5SM-splicing cassette (boxed) within gene constructs TFIIIA-eGFP (native context, top) and eGFP-P5SME/R (foreign context, bottom). The full sequence maps for these constructs are shown in Supplementary Figure S6. The eGFP coding sequence is shaded. The cassette exon contains the P5SM RNA element and an in-frame premature termination codon (X). The splicing reactions that generate spliced products SP-I and SP-II are shown. (B) eGFP-P5SME/R spliced products detected by RT–PCR upon induction with AtL5 (+) or LUC as a control (−). (C) Relative amounts of the spliced products of eGFP-P5SME/R determined by RT–qPCR. Data are averaged with number of biological replicates (n) and standard deviations shown. The p-value was determined using the paired t-test. (D) Fold induction of protein expression quantified by eGFP fluorescence for each construct. Induction with AtL5 was measured in comparison to induction with LUC as a control. (E) Representative whole leaf scan of eGFP fluorescence 3 days post-infiltration for each construct. The right leaf halves coexpressed AtL5 and the left halves coexpressed LUC (control).