棉花学报2012，Vol.24Issue(5)：468-472,5.
棉花叶片蛋白质组双向电泳技术的优化
Optimization of Two-Dimensional Electrophoresis for Cotton Leaf Proteomics
摘要
Abstract
Using cotton(Gossypium hirsutum L.) leaves as experimental materials, the present study aimed at solving the problem of disturbance by secondary metabolites in the materials, such as chromatophores, phenols and quinones, and optimizing the protein extraction method, lysis buffer reagent, protein loading amounts and other factors. The results showed that optimal cotton leaf protein extraction was obtained by the improved TCA/acetone method, with lysis buffer containing 7 mol·L-1 Urea, 2 mol-L-1 Thiourea, 4% CHAPS, 80 mmol·L-1 DTT, 1 mmol·L-1 PMSF, and 0.3% Bio-Lyte, a protein loading amount of 600 μg, and an IPG (pH 4-7, 24 cm). Under these conditions a good protein pattern with protein spots distributed equally, of high quality and repeatability, was obtained. This system will establish a foundation for cotton proteomic research.关键词
蛋白质组/双向电泳/棉花/叶片Key words
proteome/ two-dimensional electrophoresis/ cotton/ leaf分类
农业科技引用本文复制引用
郭忠军,陈全家,许春华,王希东,苏秀娟,曲延英..棉花叶片蛋白质组双向电泳技术的优化[J].棉花学报,2012,24(5):468-472,5.基金项目
国家自然科学基金(30960201) （30960201）
新疆维吾尔自治区自然科学基金(2010211A16) （2010211A16）
