Comparison of 2-DE Techniques for Improved Proteomic Analysis of Fruit Tissues
果实蛋白质组学研究的实验方法

Fruit tissues are considered recalcitrant plant tissue for proteomic analysis. In this study, several 2-D electrophores is (2-DE) protocols were evaluated to establish a reference map for the fruit proteome. Three protein extraction procedures for 2-DE and different electrophoresis systems of isoelectric focusing (IEF), as well as staining protocols, were compared with several fruit proteins amples. We achieved satisfactory 2-DE patterns with well-resolved polypeptide spots throughout the gel by homogenization (Homo) and phenol (Phe) protein extraction protocols in combination with a lysis buffer of 7 mol.L-1 urea, 2 mol.L-1 thiourea, 4% (v /v) CHAPS, 1% (w /v) DTT, 0.2% (v/v) Nonidet P40, and 2% (v/v ) carrier ampholytes. As well, immobilized pH gradient ( IPG)-IEF produced high-quality 2-DE gels that were free of smearing and streaking, and all staining protocols were suitable for protein visualization.These results suggest that the Homo and Phe protocols are highly effective with fruit tissues and that the combination IPG-IEF provides satisfactory 2-DE-based proteomic analysis of fruit tisues, which is very useful for deep investigation of biochemical changes in harvested fruit by a proteomic approach.