Extraction of proteins for two-dimensional gel electrophoresis and proteomic analysis from an endophytic fungus

Abstract

_Piriformospora_ _indica_ is a mycorrhiza like fungus of the order Sebacinales,which colonizes root and forms symbiosis with almost all terrestrial plants on earth.Information about the _P._ _indica_ quantitative proteomics is limited.This limits our understanding of its multitrophic interaction with plants.A quantitative proteomic analysis of the fungus vis a vis its interaction with rhizospheric bacteria and the plant can re ect on the underlying proteins/enzymes.We describe a protocol for e cient extraction of cellular proteins from the fungus for two-dimensional gel electrophoresis following interaction with _Azotobacter_ _chroococcum_ in axenic culture.Polysaccharides and abundant salts present in Hill and Kaefer broth hinders extraction of protein from fungal biomass.Good quantity and quality of proteins are required for two-dimensional gel electrophoresis and proteomic analysis.We could use the extracts for proteomic analysis following a high resolution of 2-DE reference map of the fungus after interaction with _A.chroococcum_.