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Recombinant protein production F, yps1 , F- ypt7 and yps1 - ypt7 strains from YPD plates were selected to inoculate into 3 ml BMGY (1% yeast extract, 2% peptone, 1.34% YNB, 0.00004% biotin, 1% glycerol, 100 mM potassium phosphate, pH 6.0) at 30C and shaken at 250 rpm