水稻抗病蛋白质XA21的免疫纯化及双向电泳分离研究

doi:10.7668/hbnxb.2008.05.026

华北农学报 ›› 2008, Vol. 23 ›› Issue (5): 118-121. doi: 10.7668/hbnxb.2008.05.026

所属专题： 水稻；

水稻抗病蛋白质XA21的免疫纯化及双向电泳分离研究

白辉^1,2, 武鹏程¹, 刘丽娟¹, 李莉云^1,2

1. 河北农业大学, 生命科学学院, 河北, 保定, 071001;
2. 中国科学院, 北京基因组研究所, 北京, 101300

收稿日期:2008-03-10 出版日期:2008-10-28
通讯作者: 刘国振(1964－)，男，河北沧州人，教授，博士，博士生导师，主要从事生物化学与分子生物学研究。
作者简介:白辉(1980－)，女，河北石家庄人，博士，主要从事生物化学与分子生物学研究。
基金资助:
国家自然科学基金(30328019；30370872)

Immunoprecipitation and Two-dimensional Electrophoresis Separation of Rice Disease Resistance Protein XA21

BAI Hui^1,2, WU Peng-cheng¹, LIU Li-juan¹, LI Li-yun^1,2

1. College of Life Sciences, Hebei Agricultural University, Baoding 071001, China;
2. Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing 101300, China

Received:2008-03-10 Published:2008-10-28

摘要/Abstract

摘要： 在优化水稻叶片蛋白质提取方法的基础上，利用免疫沉淀对水稻中的XA21蛋白质进行了富集纯化，继而通过TCA-丙酮法脱盐，用尿素、硫脲溶解后进行双向电泳分离和Western检测，实现了对高分子量、低丰度表达的、可能的膜蛋白XA21的分离检测，为其翻译后修饰方式研究奠定了基础。

关键词: 水稻, 免疫沉淀, 双向电泳

Abstract: Xa21 is a broad-spectrumresistancegeneagainstricebacterial blight,whichencodesareceptor-like kinase.The separation and detection of rice expressed XA21 protein is a key step to understand Xa21-mediated disease resistance mechanism.Based on the optimization of leaf protein extraction,XA21 was enriched and purified from crude extract of rice leaf by anti-cMyc antibody immunoprecipitation.The purified XA21 was precipitated by TCA-acetone and redissolved by urea and thiourea containing solutions,then the sample was separated by two-dimensional electrophoresis and XA21 protein was detected by western.The separation and detection of high molecular weight,low abundance and predicted membrane anchor XA21 protein provides the fundamental basis of its modification characterization.

Key words: Rice, Immunoprecipitation, Two dimensional electrophoresis

中图分类号:

Q753

白辉, 武鹏程, 刘丽娟, 李莉云, 刘国振. 水稻抗病蛋白质XA21的免疫纯化及双向电泳分离研究[J]. 华北农学报, 2008, 23(5): 118-121. doi: 10.7668/hbnxb.2008.05.026.

BAI Hui, WU Peng-cheng, LIU Li-juan, LI Li-yun, LIU Guo-zhen,. Immunoprecipitation and Two-dimensional Electrophoresis Separation of Rice Disease Resistance Protein XA21[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2008, 23(5): 118-121. doi: 10.7668/hbnxb.2008.05.026.

http://www.hbnxb.net/CN/Y2008/V23/I5/118

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水稻抗病蛋白质XA21的免疫纯化及双向电泳分离研究

Immunoprecipitation and Two-dimensional Electrophoresis Separation of Rice Disease Resistance Protein XA21

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水稻抗病蛋白质XA21的免疫纯化及双向电泳分离研究

Immunoprecipitation and Two-dimensional Electrophoresis Separation of Rice Disease Resistance Protein XA21

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