欢迎访问《中国农学通报》,

中国农学通报 ›› 2010, Vol. 26 ›› Issue (13): 279-283.

所属专题: 生物技术 园艺

• 林学 园艺 园林 • 上一篇    下一篇

叶用莴苣叶片蛋白质双向电泳技术体系的建立

张侨 韩莹琰 范双喜 陈青君 陈学珍   

  • 收稿日期:2009-12-16 修回日期:2010-03-10 出版日期:2010-07-05 发布日期:2010-07-05

Establishment of technique of proteome two-dimensional electrophoresis of the leaf of Lactuca sative L.

  • Received:2009-12-16 Revised:2010-03-10 Online:2010-07-05 Published:2010-07-05

摘要:

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,比较了两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。

关键词: 大丽花, 大丽花, 引种, 露地栽培

Abstract:

In order to establish a two-dimensional gel electrophoresis (2-DE) protocol for proteomic study of Lactuca sative L., with leaves of Lactuca sative 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 sative L.was successfully established.