Figure 2.

Analysis of the fate of the larch mitochondrial tRNA^His precursor transcript having no G_–1 in potato mitochondria. (A) Schematic representation of the strategy. First, G_–1 and G_–2 encoded by the larch mitochondrial trnH gene are deleted. Then, upon DNA uptake into potato mitochondria, larch mitochondrial tRNA^His is transcribed from the potato 18S rRNA promoter sequence (gray box). Total nucleic acids were analyzed by cRT-PCR. (B) Image of the ethidium bromide-stained gel of PCR product amplified using primers P3 and P4. The presence (+RT) or absence (–RT) of reverse transcriptase during the cDNA synthesis in the presence of primer P3 is indicated. The lane marked M shows the migration of the DNA ladder. (C) The 70-bp PCR product shown in (B) was cloned and 21 clones were sequenced. A sequence showing the junction (black vertical arrow) between 5′- and 3′- termini is presented. This sequence shows that the CCA triplet and the G_–1 (underlined) have been post-transcriptionally added.