pHIL‐S1‐opt‐RABV‐G

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	Recombinant P. pastoris clones were grown in YPD agar plates or RDB agar plates at 30°C. For expression studies, 2 ml of BMGY in culture plate (Dominique Dutscher, Brumath, France) were grown overnight at 30°C and 250 r.p.m. After 14–16 h, optical density was measured at 600 nm, and cells were resuspended in 2 ml of fresh BMMY medium to an initial OD600 of 1.

Protocol tips
Recombinant P. pastoris clones were grown in YPD agar plates or RDB agar plates at 30°C. For expression studies, 2 ml of BMGY in culture plate (Dominique Dutscher, Brumath, France) were grown overnight at 30°C and 250 r.p.m. After 14–16 h, optical density was measured at 600 nm, and cells were resuspended in 2 ml of fresh BMMY medium to an initial OD600 of 1.

Publication protocol

Recombinant P. pastoris clones were grown in YPD agar plates or RDB agar plates at 30°C. For expression studies, 2 ml of BMGY in culture plate (Dominique Dutscher, Brumath, France) were grown overnight at 30°C and 250 r.p.m. After 14–16 h, optical density was measured at 600 nm, and cells were resuspended in 2 ml of fresh BMMY medium to an initial OD600 of 1. Cultures were performed at 250 r.p.m., 30°C up to 72 h with methanol added to 1% every 24 h. Aliquots were taken every 24 h, cells were pelleted; supernatants and cells were stored at −20°C for further analysis.

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