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适合水稻悬浮培养细胞蛋白质组分析的双向电泳技术 被引量:2

A Two-dimensional Gel Electrophoresis Protocol Suitable for Proteomic Study of Rice Suspension Cells
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摘要 水稻是研究植物蛋白质组学的一个重要试材,从蛋白质的提取,裂解缓冲液的成份和浓度,第一向聚焦的条件,蛋白上样量和二向胶的选择方面,对水稻悬浮培养细胞的双向电泳条件进行了比较和优化。结果表明,采用甲醇/氯仿沉淀蛋白,裂解缓冲液中加入0.5%的两性电解质、并加入盐桥,17cm的胶条、850μg的上样量可达到较好的等电聚焦效果;8%-16%梯度胶分离蛋白较为适宜。 Rice(Oryza sativa)is a widely used material in studying plant proteomics.We optomized sample preparation method,reagent and concentration of lysis buffer,focusing condition,loading amounts of protein and the SDS-PAGE condition.The results showed that methanol/cholroform method,lysis buffer contained 0.5% ampholine,paper bridge and loading quantity of sample protein 850 μg/(17 cm IPG strip)could obtain better focusing effect;8%-16% gradient gel was appropriate for protein seperation.
出处 《生物技术通报》 CAS CSCD 北大核心 2010年第11期81-86,共6页 Biotechnology Bulletin
基金 北京市科技新星计划项目(2008B22) 北京市属市管高校人才强教计划资助项目 北京市教委项目(KM200610020005)
关键词 水稻 悬浮培养细胞 双向电泳 Rice Suspension cells Two-dimensional gel electrophoresis
作者简介 刘悦萍,女,副教授,研究方向:果实发育生理及分子生物学;E-mail:cauping@sina.com
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