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Codon Optimization | … Mutagenesis and protein expression The synthesized genes of ABL3, ABL4 and SAMEA51 codon- optimized to E coli (GenScript, USA) were incorporated into pET21a(+) vector within N-terminal NdeI and C-terminal XhoI restriction sites (Merck KGaA, Germany) … | Get A Quote |
Definition of rhodopsin is the retinal-binding membrane protein with the Schiff base linkage at a lysine on the 7th transmembrane helix. However, ~ 600 microbial rhodopsins lack retinal-binding lysine at the corresponding position (Rh-noK) among ~ 5500 known microbial rhodopsins, suggesting that Rh-noK has each functional role without chromophore. Here, we report successful functional recovery of Rh-noK. Two Rh-noKs from bacteria were heterologously expressed in Escherichia coli, which exhibited no color. When retinal-binding lysine was introduced, one of them gained visible color. Additional mutation of the Schiff base counterion further gained proton-pumping activity. Successful engineered functional recove... More