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. Author manuscript; available in PMC: 2009 Oct 21.
Published in final edited form as: Mol Genet Metab. 2008 Jan 24;94(1):68–77. doi: 10.1016/j.ymgme.2007.11.019

Figure 3. Assay of functional N-terminal leader sequences.

Figure 3

A. Schematic of the different mitochondrial leader constructs made from the pAcGFP1-mito vector. 1. Original pAcGFP1-vector with cytochrome C oxidase subunit VIII leader sequence (positive control). 2. Cytochrome C oxidase subunit VIII leader sequence is cut out, leaving the vector with expression of GFP but no mitochondrial leader (negative control). 3. pAcGFP with the 30bp 5′UTR and first 173 base pairs (47 amino acids) of MSR variant 2 attached to the N-terminus of GFP. 4. The first coding 183 base pairs (61 amino acids) of MUT attached to the N-terminus of GFP. 5. The first coding 261 base pairs (87 amino acids) of MMAB attached to the N-terminus of GFP. B. Brightfield fluorescent images of the constructs described and numbered as above, after transfection into MCH65 fibroblasts. Note the punctate pattern of GFP expression in panels 2, 4 and 5 and diffuse expression in panels 1 and 3.