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. 2018 Mar 21;15(6):726–738. doi: 10.1080/15476286.2018.1445404

Figure 2.

Figure 2.

The tRNA-like sequence regulates the splicing of its host intron and the biogenesis of the GUT15-tRF-F5 small RNA. (A) Schematic representation of the GUT15 gene variants used. Black boxes represent exons, and lines depict introns. Positions of the tRNA-like and GUT15-tRF-F5 sequences are marked in blue and red, respectively. NOS, transcription terminator. (B) Primary transcripts and splicing isoforms of GUT15 detected by RT-PCR in N. benthamiana leaves infiltrated with the GUT15 gene variants shown in A. The expression of the hygromycin phosphotransferase gene (HptII) served as a positive control for the agroinfiltration experiment. Marker, 100 bp plus (Thermo Fisher Scientific). MOCK is a negative control for the agroinfiltration experiments (leaves infiltrated only with MES buffer). (C) The accumulation levels of the tRNA-like and GUT15-tRF-F5 sequences detected by Northern blotting in infiltrated leaves expressing the GUT15 gene variants shown in A. U6 snRNA serves as an RNA loading control. Marker, Decade™ Marker RNA (Thermo Fisher Scientific).