In vivo import of unspliced tRNATyr containing synthetic introns of variable length into mitochondria of Leishmania tarentolae

摘要

A splicing-deficient tRNATyr was used as a vehicle to introduce foreign sequences into the mitochondrion of Leishmania tarentolae [Sauroleishmania tarentolae]. The naturally occurring intron was replaced by synthetic sequences of increasing length and the resulting tRNATyr precursors were expressed in transgenic cell lines. Whereas stable expression of precursor tRNATyr was obtained for introns up to a length of 76 nt, only precursors having introns up to 38 nt were imported into mitochondria. It is concluded that splicing-deficient tRNATyr can be used to introduce short synthetic sequences into mitochondria in vivo, and one factor which limits the efficiency of import is the length of the molecule.