茭白总蛋白质双向电泳技术体系的建立

为建立茭白总蛋白质双向电泳技术体系,研究了TCA/丙酮沉淀法和改良酚抽法两种不同总蛋白质提取方法、蛋白质上样量、pH值范围及凝胶质量浓度等条件对2-DE的影响。结果表明,改良酚抽法更适合茭白总蛋白质的提取,采用17 cm、pH 4~7的胶条、1.0 mg的蛋白质上样量、12 g/dL的凝胶浓度、考马斯亮蓝G-250胶体考染法染色,最终可获得蛋白质点较多,背景清晰,分辨率较高的2-DE图谱,为进一步开展茭白差异蛋白质组学研究奠定了基础。

Establishment of Two-Dimensional Electrophoresis Conditions for Proteomic Analysis of Zizania latifolia

In order to establish the two-dimensional electrophoresis(2-DE) conditions of Zizania latifolia,the effects of different protein extraction methods,loading amount,pH ranges of IPG strip and gel concentration,etc.on 2-DE maps were investigated.The results showed that the modified phenol extraction protocol is more suitable for total protein extraction of Z.latifolia compared with TCA/acetone protocol.And a high quality 2-DE map with more protein spots,clear background and high protein point resolution was obtained using the following optimized procedure:loading 1.0mg protein sample on 17 cm IPG strip with pH 4-7,SDS-PAGE with 12 g/dL gel concentration,and finally detecting proteins with coomassie brilliant blue G250 staining.The research provides a basis for further study of the Z.latifolia proteomics.