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Medicine and Health Sciences

University of the Pacific Theses and Dissertations

2009

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Comparison Of Zeocin And G418 Resistance Markers In Pichia Pastoris, Allison Moy Jan 2009

Comparison Of Zeocin And G418 Resistance Markers In Pichia Pastoris, Allison Moy

University of the Pacific Theses and Dissertations

The methylotrophic yeast, Pichia pastoris, has been used to successfully produce and express over 700 heterologous proteins. Currently, the only two dominant selectable markers available are the Zeocin and blasticidin resistance genes. This study focused on characterizing and developing the modified G418 selectable marker so it could be used for primary selection of single copy and multicopy strains. To demonstrate its use as a selectable marker the SLPI reporter gene was inserted into the Zeocin resistance vector, pPICZB, and the new G418 resistance vector, pKANa B. The resulting constructs were then transformed into P. pastoris yJC100 cells. Piggyback strains containing …