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Recombinant pPinkα-appA plasmid was linearized by digestion with BspT1 restriction enzyme and it was transformed into P. pastoris GS115 by electroporation (BTX, ECM630 at 1800 V, 200 Ω and 25 µF with 0.2 cm cuvette), following the Invitrogen protocol. After the transformation, the cuvette contents were spread on PAD plates (Pichia Adenine Dropout) and incubated at 30°C for 10 days. As a negative control, the vector of pPinkα-HC was transformed into P. pastoris. |
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| Protocol tips |
| Recombinant pPinkα-appA plasmid was linearized by digestion with BspT1 restriction enzyme and it was transformed into P. pastoris GS115 by electroporation (BTX, ECM630 at 1800 V, 200 Ω and 25 µF with 0.2 cm cuvette), following the Invitrogen protocol. After the transformation, the cuvette contents were spread on PAD plates (Pichia Adenine Dropout) and incubated at 30°C for 10 days. As a negative control, the vector of pPinkα-HC was transformed into P. pastoris. |
Publication protocol
Recombinant pPinkα-appA plasmid was linearized by digestion with BspT1 restriction enzyme and it was transformed into P. pastoris GS115 by electroporation (BTX, ECM630 at 1800 V, 200 Ω and 25 µF with 0.2 cm cuvette), following the Invitrogen protocol. After the transformation, the cuvette contents were spread on PAD plates (Pichia Adenine Dropout) and incubated at 30°C for 10 days. As a negative control, the vector of pPinkα-HC was transformed into P. pastoris.
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