Heterologous expression of secreted proteins from Phytophthora in Pichia pastoris followed by protein purification by Immobilized Metal Ion Affinity (IMAC)

Abstract

This chapter describes the protocol for heterologous expression of Phytophthora proteins in the yeast Pichia pastoris. Two methods to prepare the constructs for expression are described, using two different strains of P. pastoris, as well as methods for protein expression and purification by immobilized metal ion affinity (IMAC). The combined protocol was used to successfully produce a Phytophthora pluvialis small, secreted cell death elicitor protein and a Phytophthora agathidicida glycoside hydrolase 12, PaXEG1, also a cell death-inducing protein