叶用莴苣叶片蛋白质双向电泳技术体系的建立.pdf

中国农学通报 2010,26(13):279-283
Chinese Agricultural Science Bulletin
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
(北京农学院植物科学技术学院,北京 102206)
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 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。
关键词:叶用莴苣;蛋白提取;双向电泳
中图分类号: 文献标志码:A 论文编号:2009-2677
Establishment of Technique of Proteome two-dimensional Electrophoresis of
the Leaf of Lactuca sative L
Zhang Qiao, Han Yingyan, Fan Shuangxi, Chen Qingjun, Chen Xuezhen
( , , Beijing 102206)
Department of Plant Science and Technology Beijing Agricultural College
In order to establish a two-dimensional gel electrophoresis (2-DE) protocol for proteomic study of
Abstract:
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 an