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学科主题: 物理化学
题名: A new method for identification the oligochitosan binding protein on tobacco plasma membrane
作者: Yin H(尹恒) ;  Du YG(杜昱光)
会议名称: Annual Main Meeting of the Society of Experimental Biology
会议日期: 2009-6-28
出版日期: 2009-06-28
会议地点: 英国
其他题名: 一种鉴定壳寡糖与烟草质膜蛋白结合的新方法
通讯作者: 尹恒
部门归属: 十八室
主办者: 实验生物学会
摘要: Oligochitosan is a potent plant immunity elicitor, however, themechanism on howoligochitosan induces the resistance of plants to pathogens is still unclear. To understand the molecular mechanism regarding how plant cells perceive the oligochitosan signal, identifying the receptors for the elicitors is very important. In this work, a convenient method for identification of the oligochitosan binding protein on tobacco plasma membrane was found. Oliogochitosan labeled with the green fluorescent probe 2-aminoacidone (2-AMAC) was prepared and 2-AMAC was used as negative control. Tobacco plasma membrane was purified by using aqueous twophase partitioning and separated by polyacrylamide gel electrophoresis. After electrophoresis, the gel was washed by Triton-100 several times to renatured the protein function. Then the gel was incubated with 2-AMAC-oligochitosan at 25 °C for 30 min and the unincorporated 2-AMAC-oligochitosan was removed by washing in the PBS buffer for at least 6 h with five changes. Binding of oligochitosan with tobacco plasma membrane was observed under ultraviolet detector. The bands which presented green fluorescence were recovered from the gel with ordinary electrophoresis apparatus and sequenced by MALDI-TOF MS. This method is quick and non-radioactive for carbohydrate–protein interaction and identification compared to general isotope labeling method.
语种: 中文
Citation statistics: 
内容类型: 会议论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/113586
Appears in Collections:中国科学院大连化学物理研究所_会议论文

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Recommended Citation:
Yin H,Du YG. A new method for identification the oligochitosan binding protein on tobacco plasma membrane[C]. 见:Annual Main Meeting of the Society of Experimental Biology. 英国. 2009-6-28.
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