Figure 5.

Schematic representation of the intron constructs used to examine the splicing of the tRNA/tRNA-like carrying introns. (A) Schematic structure of the pDH515 vector with marked positions of the inserted intron constructs. Closed box, the zein gene; open boxes, the CaMV 35S promoter and terminator regions. (B) Schematic diagrams of the intron constructs used. Boxes represent exons, and lines depict introns. The sizes of the original exon fragments used in the constructs are shown. (C) RT-qPCR analysis of the splicing efficiency of particular intron constructs. Arrows shown in the upper part of the panel with the intron mini-construct schemes depict the primers used. Intron retention isoform levels were calculated as a percentage of all splicing events (intron retained (IR) plus fully spliced (FS) transcripts, treated as 100%) identified within the analyzed intron. Error bars indicate SD (n = 3), and the asterisk indicates a significant difference in the splicing efficiency between the native construct and the mutated constructs (*p < 0.05).