Protocols for nuclei isolation and nuclear protein extraction from the resurrection plant Xerophyta viscosa for proteomic studies

A rapid, simple in vitro test system for high-throughput screening of peroxisome proliferator-activated receptor (PPAR) γ agonists would be of interest for testing new antidiabetic drugs, alternative medicine, or environmental samples. A yeast two-hybrid assay based on the ligand-dependent recruitment of the coactivator CBP (CREB-binding protein) was constructed. In this system PPARγ was constitutively activated and the signal was not further increased significantly by adding agonists. In yeast we identified oleic acid as a putative endogenous ligand. Furthermore yeasts seem to lack regulatory mechanisms present in mammalian cells. Mammalian systems are an alternative for screening PPARγ agonists.