Part:BBa_K210010:Experience

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Applications of BBa_K210010

iGEM Kyoto 2009

To confirm the function of the signal sequence for importing protein into mitochondria, we compared the expressioin pattern of sig-GFP or GFP with mitotracker signal. The GFP signal was detected throughout the cell except for the black granules in the cytoplasm or the nuclear, while sig-GFP signal showed the string-like pattern in the cytoplasm. The black granules in the cytoplasm observed in the GFP-expressing cell were stained by mitotracker, and the result indicated that the GFP was normally eliminated from mitochondria (Fig.1, GFP and mitotracker merged). In case of sig-GFP, mitochondria stained by a mitotracker and the GFP signal showed almost the same pattern. the yellow color in the merge images (Fig. 2, sig-GFP and mitotracker merged) suggested that the sig-GFP and mitochondria were colocalized in the HeLa cell. We, consequently, our constructed signal sequence has the function of importing protein into mitochondria as expected.

Figure 1: Conforcal microscopic images of GFP or sig-GFP transfected cells. The image lines titled "mitotracker(+)" indicated the samples was stained by mitotracker. The columns showed the mitotracker, GFP, and merged images from left, respectively.

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