Figure 1.

Construction and expression of the mitochondrially targeted EcoRI. A DNA fragment coding for the COX VIII targeting sequence (MTS) was added to the 5′ end of the restriction endonuclease gene EcoRI and as optical marker the gene for the enhanced green fluorescent protein EGFP was fused to the 3′ end. The construct was sub-cloned into the vector pTRE2hyg and re-cloned with AgeI and NotI into pEGFP-Mito resulting in the vector pMEE-con with a constitutive CMV promoter (A) The final construct was transfected in 143B.DsRed1-Mito cells. Twenty-four hours after transfection the mitochondrial localization of the EcoRI fusion protein in punctate structures (white arrowheads) was confirmed by co-localization with DsRed1-Mito with confocal fluorescence microscopy (B) The calibration mark corresponds to 10 µm.