KpTrm1p

Protein Expression Eukaryotic cells - P. pastoris phytase

Experiment
Protein Expression Eukaryotic cells - P. pastoris phytase
Product
KpTrm1p from Yasuyoshi Sakai, Division of Applied Life Sciences, Graduate Sch
Manufacturer
Yasuyoshi Sakai, Division of Applied Life Sciences, Graduate Sch

Protocol tips

Protocol tips
K. phaffii strains were transformed by electroporation following the manufacturer's protocol (Invitrogen, Cat. No. K1710–01).

Publication protocol

K. phaffii strains were transformed by electroporation following the manufacturer's protocol (Invitrogen, Cat. No. K1710–01). RD medium (1 M sorbitol, 2% dextrose, 1.34% yeast nitrogen base (YNB), 4 × 10−5% biotin, 0.005% each amino acid of L-glutamic acid, L-methionine, L-lysine, L-leucine, L-isoleucine) supplemented with 2% Difco Agar Noble or MD medium (2% dextrose, 1.34% YNB, 4 × 10−5% biotin) supplemented with 1 M sorbitol and 2% agar noble was used for regeneration with his4 selection. In the case of Zeocin resistance selection, YPDS + Zeocin agar (1 M sorbitol, 2% dextrose, 1% yeast extract, 2% peptone, 2% bacto agar, 100 μg/mL Zeocin (Life Technologies, Tokyo, Japan)) was used for the regeneration. Plates were incubated at 28 or 30°C for 3–4 days until colonies appeared. Screening for Mut+/MutS phenotypes was performed using MD (minimum dextrose) medium plates and MM (minimum methanol) medium plates following the published protocol (Invitrogen Catalog Number K1710–01).

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