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以烟草品种K326旺长后期叶片为材料,用IEF/SDS-PAGE双向凝胶电泳技术对5种蛋白质提取方法(TCA/丙酮沉淀法、Mg/NP-40法、苯酚法、TCA-苯酚法和可溶性蛋白质提取法)进行比较,并对自制管胶pH范围进行探索。结果表明:苯酚法图谱中蛋白质条带最多、最清晰,并可获得较高纯蛋白质得率,为0.0420%,且图谱中蛋白质条带数最多、最清晰。自制管胶pH范围优化试验结果表明,以两性电解质pH 3-10与pH 5-8比例为1:3的2-DE图谱质量最佳,蛋白质可得到较好的分离。
The leaves of tobacco variety K326 Wangchang were used as material, and five protein extraction methods (TCA / acetone precipitation, Mg / NP-40 method, phenol method, TCA- phenol method and IEF / SDS-PAGE two-dimensional gel electrophoresis Soluble protein extraction method) to compare, and to explore the pH range of homemade tube glue. The results showed that the bands of phenol in the phenol method were the most and the most clear, and the yield of pure protein was 0.0420%, and the number of protein bands was the most and the most clear. The results of the optimization of the pH range of the self-made tube showed that the 2-DE map with the ratio of amphoteric electrolyte pH 3-10 to pH 5-8 of 1: 3 was the best, and the protein was well separated.