pET-PvD1

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	The transformed cells were grown at 37°C in liquid LB medium (0.5% yeast extract, 1.0% tryptone, 1.0% NaCl) supplemented with ampicillin (50 μg.mL-1) and chloramphenicol (34 μg.mL-1) to an optical density (at 600 nm) of 0.5 before induction with 1 mM IPTG.

Protocol tips
The transformed cells were grown at 37°C in liquid LB medium (0.5% yeast extract, 1.0% tryptone, 1.0% NaCl) supplemented with ampicillin (50 μg.mL-1) and chloramphenicol (34 μg.mL-1) to an optical density (at 600 nm) of 0.5 before induction with 1 mM IPTG.

Publication protocol

The transformed cells were grown at 37°C in liquid LB medium (0.5% yeast extract, 1.0% tryptone, 1.0% NaCl) supplemented with ampicillin (50 μg.mL-1) and chloramphenicol (34 μg.mL-1) to an optical density (at 600 nm) of 0.5 before induction with 1 mM IPTG. After 3 h of induction, the cells were harvested by centrifugation (4,000 g for 15 min at 4°C); suspended in 50 mM phosphate buffer (pH 7.4) containing 500 mM NaCl, 40 mM imidazole, 1 mM PMSF, and a protease inhibitor cocktail for general use (Sigma, USA), according to the manufacturer’s manual; and ruptured by sonication (10 pulses of 30 sec at 10 watts) (R2D91109, Unique). Next, the cell lysate was clarified by centrifugation (5,000 g for 10 min at 4°C) (5430R, Eppendorf), and the supernatant was heated at 90°C for 30 min. The resulting suspension was centrifuged (7,200 g for 10 min at 4°C), and the supernatant was purified further by chromatographic methods.

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