缺铁逆境胁迫下水稻叶蛋白质组的双向电泳分析  

2-DE Analysis of Differentially Displayed Proteins in Rice Young Shoots Under Iron-deficiency Stress

作　　者：柴小清[1] 靳飞[1] 张艳萍[1] 卫功宏[2] 印莉萍[1] 

机构地区：[1]首都师范大学生物系,北京100037 [2]中国协和医科大学基础医学院,北京100005

出　　处：《首都师范大学学报（自然科学版）》2004年第3期46-51,共6页Journal of Capital Normal University：Natural Science Edition

摘　　要：水稻幼苗经缺铁胁迫诱导处理 1、3、5d后 ,用酚法和TCA 丙酮法提取叶的可溶性蛋白 ,进行双向电泳 (2 DE)分析 .结果显示 :(1 )三种缺铁胁迫的可溶性蛋白 ,在不同的pH范围中 2 DE图谱上的比较 .使用pH3～ 1 0宽范围、线性 (L)的IPG胶条 ,经电泳分离后利用Z3图象分析软件 ,可在SDS PAGE凝胶上检测到 4 5 0个左右的蛋白点 ,酸性蛋白占 89% .选用pH4～ 7窄范围、线性 (L)的IPG胶条 ,经电泳后可在SDS PAGE凝胶上检测到 6 0 0个左右的蛋白点 ,其中缺铁诱导上调表达的有 2 9个点 ,减弱表达的有 1个点 ,诱导特异表达的有 5个点 .(2 )不同方法提取可溶性蛋白的差异 .酚法提取的蛋白再溶性好 ,纯度较高 .TCA 丙酮法简单易操作 ,蛋白损耗少 ,在 2 DE图象上 ,碱性端显示的蛋白点较多 ,但此法的缺点是再溶性差 ,对 2Employing high resolution two dimensional gel electrophoresis(2 DE), the differentially displayed proteins expressed in rice( Oryza sativa ) leaves under iron deficiency stress were separated. The soluble proteins of rice leaves were extracted by phenolic method and TCA strategy respectively. Rice seedlings were grown 1,3,5d in the absence of Fe separately. The results shows below.(1) Differentially separative effect were obtained by using different pH gradient IPG gel stripe. When the proteins were visualized with colloidal Coomassie blue and analyzed by Z3 image software we detected nearly 450 protein spots on the 2 DE gels with a pH 3～10 gradient IPG stripe. Among these spots acid proteins' rate was about 89%. In addition, with a pH 4～7 gradient IPG stripe we found about 600 spots on the 2 DE gels. 29 proteins were up regulated, only 1 spot were down regulated, about 5 proteins were differentially displayed. (2) The quality and quantity of the soluble proteins were different with different extraction methods. The proteins which were extracted by phenolic method were highly quantitative and pure. TCA technique was easy to operate. It seemed very effective to basic proteins. A large quantity of protein spots appeared on the basic apex of 2 DE gels. However, the proteins which were extracted by this method can't resolubilized thoroughly.

关 键 词：可溶性蛋白 2-DE 白质 诱导 缺铁胁迫 表达 双向电泳分析 叶蛋白 水稻 SDS-PAGE 

分 类 号：N94[自然科学总论—系统科学] S511[农业科学—作物学]