pASKMscL-EGFP-SuptoxD

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	E. coli cells freshly transformed with the 465 appropriate expression vector(s) were used for all protein production experiments.

Protocol tips
E. coli cells freshly transformed with the 465 appropriate expression vector(s) were used for all protein production experiments.

Publication protocol

E. coli cells freshly transformed with the 465 appropriate expression vector(s) were used for all protein production experiments. 466 Single bacterial colonies were used to inoculate liquid LB cultures containing the 467 appropriate combination of antibiotics (100 たg/mL ampicillin, 40 たg/mL 468 chloramphenicol or 50 たg/mL kanamycin (Sigma)). These cultures were used with a 469 1:50 dilution to inoculate fresh LB cultures with 0.01% (MC1061 and SuptoxD) or 470 0.2% arabinose (SuptoxR), which were grown at 30 °C to an optical density at 600 nm 471 (OD600) of 漢0.3−0.5 with shaking, unless specified otherwise. The temperature was 472 then decreased to 25 °C and after a temperature equilibration period of 10−20 min, MP 473 expression was induced by the addition of 0.2 たg/mL aTc (Sigma) overnight, unless 474 specified otherwise. For rhodopsin overproduction, we followed the same procedure, 475 but when the cell density reached OD600 漢0.3−0.5, protein production was induced by 476 the addition of 0.2 たg/mL aTc in the presence of 10 たM all-trans-retinal (Cayman 477 Chemical) overnight in dark.

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