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Yam chitinase E
Vacuolar targeting signal (VTS)
AOX 1 AOX 1 TT
a-factor
ZeocinR
AOX 1 AOX 1 TT pPICZaA / YamE without VTS
Multiple cloning site
pPICZaA
ZeocinR
a-factor Yam chitinase E with VTS
Ligation
Yam chitinase E
AOX 1 AOX 1 TT
Transformation of Eschericia coli JM109 and grown on LB medium containing ZeocinTM(at 37℃)
Plasmid extraction
Fig. 4. Outline of Yam class IV chitinase expression systems in P. pastoris. The vector
pBluescript SK+ and pDrive were used respectively for VTS and no-VTS yam chitinase cloning.