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pAM1 (AOX1:SLPI), pCH3K(AOX1:HRP), pKanB-lipase (AOX1:lipase) and pKanB (control) were linearized with MssI or Mph1103I and transformed into electrocompetent mutant (bgs2, bgs3, bgs13) or wild type cells (yDT39:pBLMET-βgal) and selected on medium containing 500 µg G418/ml (Lin-Cereghino et al. 2013). pHSA413 was linearized with SalI and transformants were selected on YND. Colony PCR (Thor et al. 2005) was performed to confirm that transformants harbored the reporter genes. |
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| Protocol tips |
| pAM1 (AOX1:SLPI), pCH3K(AOX1:HRP), pKanB-lipase (AOX1:lipase) and pKanB (control) were linearized with MssI or Mph1103I and transformed into electrocompetent mutant (bgs2, bgs3, bgs13) or wild type cells (yDT39:pBLMET-βgal) and selected on medium containing 500 µg G418/ml (Lin-Cereghino et al. 2013). pHSA413 was linearized with SalI and transformants were selected on YND. Colony PCR (Thor et al. 2005) was performed to confirm that transformants harbored the reporter genes. |
Publication protocol
pAM1 (AOX1:SLPI), pCH3K(AOX1:HRP), pKanB-lipase (AOX1:lipase) and pKanB (control) were linearized with MssI or Mph1103I and transformed into electrocompetent mutant (bgs2, bgs3, bgs13) or wild type cells (yDT39:pBLMET-βgal) and selected on medium containing 500 µg G418/ml (Lin-Cereghino et al. 2013). pHSA413 was linearized with SalI and transformants were selected on YND. Colony PCR (Thor et al. 2005) was performed to confirm that transformants harbored the reporter genes.
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