叶用莴苣叶片蛋白质双向电泳技术体系的建立

Establishment of Technique of Proteome two-dimensional Electrophoresis ofthe Leaf of Lactuca sative L

为建立适用于生菜的(Lactuca sative L)蛋白质组研究的双向电泳技术(2-DE),以叶用莴苣S24为材料,采用TCA/丙酮沉淀法提取叶片蛋白,比较了2种标准曲线对蛋白浓度测定的影响,同时探讨了蛋白裂解、等电聚焦(IEF)条件以及其他一些关键技术,结果显示:与传统标准曲线相比,采用改良后标准曲线测定的蛋白浓度更准确,得到了背景清晰、重复性好、蛋白点分辨率较高的双向电泳图谱。初步建立了叶用莴苣叶片蛋白质的双向电泳技术(2-DE)体系。

In order to establish a two-dimensional gel electrophoresis （2-DE） protocol for proteomic study of Lactuca satire L, with leaves of Lactuea satire L （S24） as material, we used TCA / acetone precipitation method extracted leaf protein, compared the two kinds of standard curve for protein concentration determination of the impact and also discussed on the lysis method, process of IEF and some key technologies of 2D electrophoresis.The results showed that： Compared with the traditional standard curve, using the improved standard curve more accurate determination of protein concentration. A two-dimensional electrophoresis atlas with good repeatability and high resolution was obtained. A system of two-dimensional electrophoresis of leaves of Lactuca satire L was successfully established.