| Products/Services Used | Details | Operation |
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| Codon Optimization | Dd-MCU, Af-MCU and Nc-MCU with (HsMTSAf-MCU and HsMTSNc-MCU) and without the N-terminal MTS of Hs-MCU (aminoacids 1-56) and without a stop codon were codon optimized for human expression, synthesized de novo in the PuC57 vector (GenScript) and amplified with flanked attB1 and attB2 sites by PCR using the following primers: fw-DdMCU (5 -GGG GAC AAG TTT GTA CAA AAA AGC AGG CTT AGC CAC CAT GAA CTC CTT TGT CAT CAG-3 ); rv-DdMCU (5 -GGG GAC AAG TTT GTA CAA AAA AGC AGG CTT AGC CAC CAT GAA TTG CGT GAG AAT GAG ACT C-3 ); fw-NcMCU (5 -GGG GAC AGG TTT GTA CAA AAA AGC AGG CTT AGC CAC CAT GAA TTG CGT GAG AAT GAG ACT C-3 ); rv-NcMCU (5 -GGG GAC CAC TTT GTA CAA GAA AGC TGG GTT ACT GTC TCC GCT GGT CTC TTT-3 ), fw-AfMCU (5 -GGG GAC AAG TTT GTA CAA AAA AGC AGG CTT AGC CAC CAT GGT CCT GTC TTG TGA TAC TAG A-3 ); rv-AfMCU (5 -GGG GAC CAC TTT GTA CAA GAA AGC TGG GTT GTC GTC ATC TCG GTC ATC GTT-3 ); fw-HsMTS (5 -GGG GAC AAG TTT GTA CAA AAA AGC AGG CTT AGC CAC CAT GGC GGC CGC CGC AGG TAG A-3 ). | Get A Quote |
The mitochondrial calcium uniporter is a highly selective ion channel composed of species- and tissue-specific subunits. However, the functional role of each component still remains unclear. Here, we establish a synthetic biology approach to dissect the interdependence between the pore-forming subunit MCU and the calcium-sensing regulator MICU1. Correlated evolutionary patterns across 247 eukaryotes indicate that their co-occurrence may have conferred a positive fitness advantage. We find that, while the heterologous reconstitution of MCU and EMRE in vivo in yeast enhances manganese stress, this is prevented by co-expression of MICU1. Accordingly, MICU1 deletion sensitizes human cells to manganese-dependent ce... More
