Mitochondrial Fluorescent Protein Constructs

Mito-DsRed plasmid, a gift from Dr. Quan Chen (Institute of Zoology, CAS), was constructed by fusing the mitochondrial target sequence of human TXN2 with the DsRed sequence. TOM20-GFP and mito-YFP, gifts from Dr. Li Yu (Tsinghua University)^{11 (link)}, were obtained by inserting the rat TOM20 coding sequence into pEGFP-N1 and by fusing the mitochondrial target sequence of human COX8A with the YFP sequence, respectively. We constructed the mito-BFP plasmid by replacing the DsRed of mito-DsRed with the BFP sequence. We also constructed TFAM-GFP by inserting the human TFAM coding sequence into pEGFP-N1 and TFAM-mCherry by replacing the GFP of TFAM-GFP with the mCherry sequence. TFAM-HaloTag was a gift from Dr. Dong Li (Institute of Biophysics, CAS) and mCherry-KDEL was a gift from Dr. Jianguo Chen (Peking University). We purchased mEmerald-Sec61β from Addgene (#90992). The primers used in this study are listed in Supplementary Table 1.