The mitochondrial targeting sequence was identified using the MitoProt software (http://ihg.gsf.de/ihg/mitoprot.html).html
For that purpose we added the mitochondrial targeting sequence of the PAD1 to the FDC1.
Signal sequence predicting programs identify a N-terminus putative chloroplast targeting sequence.
In both cases the cell free extract of the yeast strain over-expressing both genes, PAD1 and FDC1 with added targeting sequence or FDC1 and PAD1 with removed targeting sequence, showed no cinnamic acid decarboxylase activity.
Specific localization is achieved by fusing the FP gene to the mitochondrial targeting sequence of an endogenous protein.
The mutated EGFP coding sequence was divided into two segments, which were separated by a stop codon and targeting sequence.
The targeting sequence of gRNA was 5′-GTCGTGCACCGTGTCCTCCG-3′ with a protospacer adjacent motif (PAM) of CGG.
PINK1, a 64-KD protein, is encoded by PTEN-induced kinase1, which contains a mitochondrial targeting sequence (MTS).
The FDC1 gene however has no targeting sequence and the Fdc1p is believed to be a cytosolic protein.
Fig. 6 Subcellular localization of TCM5 protein; a, TCM5-GFP fusion; b, Empty GFP vector without a specific targeting sequence.
In contrast, the choice of the rDNA targeting sequence was found to be of impact on productivity.
