A Proteomics Sample Preparation Method for Mature,Recalcitrant Leaves of Perennial Plants |
| |
| Authors: | Deng Gang Zhong Xinyue Zhang Na Lao Chengying Wang Bo Peng Dingxiang Liu Lijun |
| |
| Institution: | 1. MOA Key Laboratory of Crop Ecophysiology and Farming System in the Middle Reaches of the Yangtze River, College of Plant Science and Technology, Huazhong Agricultural University, Wuhan, Hubei, China.; 2. School of Agricultural Science, Yunnan University, Kunming, Yunnan, China.; 3. Wuhan Agriculture and Science Institute, Huangpi District, Wuhan, Hubei, China.; Lawrence Berkeley National Laboratory, United States of America, |
| |
| Abstract: | Sample preparation is key to the success of proteomics studies. In the present study, two sample preparation methods were tested for their suitability on the mature, recalcitrant leaves of six representative perennial plants (grape, plum, pear, peach, orange, and ramie). An improved sample preparation method was obtained: Tris and Triton X-100 were added together instead of CHAPS to the lysis buffer, and a 20% TCA-water solution and 100% precooled acetone were added after the protein extraction for the further purification of protein. This method effectively eliminates nonprotein impurities and obtains a clear two-dimensional gel electrophoresis array. The method facilitates the separation of high-molecular-weight proteins and increases the resolution of low-abundance proteins. This method provides a widely applicable and economically feasible technology for the proteomic study of the mature, recalcitrant leaves of perennial plants. |
| |
| Keywords: | |
|
|
