The development of a kanamycin resistant gene for positive selection in the yeast Pichia Pastoris

The yeast Pichia pastoris is widely used as a host organism in foreign protein expression, which is a process that is utilized to produce many pharmaceuticals and other industrial products. One of the main problems of P. pastoris is that it does not have a wide range of selectable markers available for use in transformation and expression processes. This being the case, the use of P. pastoris can be expensive in that one of the few efficient selectable markers on hand is the zeocin resistance gene. Currently only one company, Invitrogen, distributes a commonly used reporter gene, the zeocin resistance gene. Zeocin is an antibiotic that binds to the DNA of a cell (e.g. P. pastoris) and cleaves it causing cell death. In this experiment we will attempt to adapt a kanamycin resistance reporter gene to fulfill the same function of the zeocin resistance reporter gene. By creating a new reporter gene using kanamycin resistance the costs of transformation in lab research will drastically be cut. With this new cassette for a reporter gene we hope to open the market for other researchers enabling them to be economically efficient in the lab