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Ion Transport Activity Assay for Microbial Rhodopsin Expressed in Cells

Bio Protoc. 2021-08; 
Masae Konno, Keiichi Inoue, Hideki Kandori
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Bacterial Expression … the accumulation of genetic information, the number of genes predicted to … for its codon for E. coli and synthesized (GenScript). The synthesized gene was cloned between the NdeI and … Get A Quote

摘要

Microbial rhodopsins have diverse functions, including roles as light-driven ion pumps, light-gated ion channels, photosensors, and light-regulated enzymes. As the number of rhodopsin-like genes identified has increased in recent years, so has the requirement for rapid identification of their functions. The patch-clamp method is often used to investigate the ion transport mechanism of microbial rhodopsins in mammalian cells; however, this requires a dedicated system and advanced techniques. The ion transport assay using the expression system described here evaluates the ion transport capacity by monitoring the pH change in suspensions; if the target rhodopsin has a light-dependent ion transport activity, a li... More

关键词

Escherichia coli, Ion transport, Light-dependent pH change, Microbial rhodopsin, Protein expression