After digestion with XhoI and XbaI, the PCR product was cloned
into the vector pPICZaA (Invitrogen).
The construct was confirmed
by DNA sequencing carried out by Macrogen and designated
pPICZaA/SBgl3His. The pPICZaA/SBgl3His plasmid was linearized
with SacI and transformed into P. pastoris GS115 by electroporation.
Positive transformants harboring the SBgl3 gene were selected
on YPDS plates containing 100 lg/ml zeocin.