Oct 29, 2010 · Mut S phenotype integrated into the grown colonies with the AOX1 locus of the yeast genome were selected and grown in 1 L of buffered glycerol complex medium to an optical density over 6 at 600 nm. Cells were induced with 1% methanol by replacing the buffered glycerol complex medium with buffered methanol complex medium.
Twenty transformants were used to inoculate 40 ml of buffered glycerol complex medium (BMGY) (Invitrogen) (0.1 M potassium phosphate buffer at pH 6.0 containing 1% [wt/vol] yeast extract, 2% [wt/vol] peptone, 1.34% [wt/vol] yeast nitrogen base without amino acids, 1% [vol/vol] glycerol, and 4 × 10 −5 % [wt/vol] biotin). Cultures were grown Then, the culture was transferred into 1 L fresh BMGY, and all of the cells were collected by centrifugation at room temperature (3,500 g for 5 min) until the of the culture reached approximately 6, at which point it was resuspended in 100 mL of buffered methanol-complex medium (BMMY). Methanol (3%) was added to the media daily to induce the Jul 27, 2017 · The cells were then recovered by adding 500 µL liquid LB medium and incubating at 37 °C for 1 h and then plated on LB plates supplemented with 25 μg/mL Zeocin (TFS, R25001). After incubated at 37 °C overnight, ten colonies were cultured in 3 mL liquid LB medium supplemented with 25 µg/mL Zeocin at 37 °C overnight. The recombinants were cultured in Buffered Glycerol- complex Medium (BMGY) at 30℃ and then resuspended in Buffered Methanol-complex Medium (BMMY). The cultures were shaken at 250 r/min for 108 h at 29℃, and the methanol concentration retained 0.5% (w/v). Column chromatography The supernatant was harvested by 3600 rpm centrifugation grown in 1 L of buffered glycerol complex medium to an optical density over 6 at 600 nm. Cells were induced with 1% methanol by replacing the buffered glycerol complex medium with buffered methanol complex medium. Methanol was added at a concentra-tion of 1% every 24 hours during the induction phase (up to 72 hours). Cultures were centrifuged All other chemicals used were analytical grade reagents unless otherwise stated. Yeast extract peptone dextrose (YPD) medium, buffered glycerol complex (BMGY) medium, and buffered methanol complex (BMMY) medium were prepared according to the manual of Pichia Expression Kit (Version F, Invitrogen). P. pastoris GS115 cells transformed with pHIL‐D2‐ACMSD I were incubated in buffered glycerol complex medium, at 30 °C, up to an A 600 of 3.0. The culture was centrifuged at 5000 g for 10 min (Sorvall centrifuge RC5B plus, Superlite GSA rotor) and the cell pellet was resuspended in one‐fifth of the original culture volume of buffered
grown in 1 L of buffered glycerol complex medium to an optical density over 6 at 600 nm. Cells were induced with 1% methanol by replacing the buffered glycerol complex medium with buffered methanol complex medium. Methanol was added at a concentra-tion of 1% every 24 hours during the induction phase (up to 72 hours). Cultures were centrifuged
Oct 15, 2018 · The protein expressing strains were obtained using G418 selection and cultured in 25 ml of buffered glycerol-complex medium [BMGY; 1% yeast extract, 2% peptone, 100 mM potassium phosphate (pH 6.0
Buffered Glycerol Complex Medium, Sterile 2.0% Peptone, 1.0% Yeast extract, 1.34% Potassiumphosphate pH 6.0, 1.34% Yeast nitrogen base w/o AA, 0.4 mg/L Biotin, 1.0%
The applications of viral protein cages have expanded rapidly into the fields of bionanotechnology and materials science. However, the low-cost produc…