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甘蓝柱头蛋白质双向电泳体系的建立 被引量：1

Establishment of Two-dimensional Electrophoresis System of Brassica oleracea L.var.capitata L.Stigma Protein
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摘要 蛋白质提取方法和电泳条件是蛋白质组学研究中双向电泳的关键。为建立适合甘蓝柱头总蛋白质的双向电泳体系,本试验以甘蓝柱头为材料,采用改良TCA/丙酮沉淀法提取柱头总蛋白质,通过对IPG胶条、上样量、聚焦程序、SDS-PAGE胶浓度等的优化,建立了甘蓝柱头总蛋白质双向电泳体系,即选用长17cm、pH3～10NL的IPG胶条,上样量150μg,按照聚焦程序Ⅰ进行等电聚焦,第二向SDS-PAGE胶浓度选用12%,得到了分辨率高、重复性好的双向电泳图片。 A two-dimensional electrophoresis(2-DE)has been widely applied in proteomics research.The purpose of this study was to establish a suitable 2-DE system for proteomic analysis of Brassica oleracea L.var.capitata L.stigma.The stigmas being harvested one day before flowering was used in the experiment.We optimized the parameters of 2-DE including extraction methods,IPG strip,sample loading,isoelectric focusing conditions and SDS-PAGE gel concentration.The optimized system included the following steps:extracting the total proteins from Brassica oleracea stigma by TCA/acetone precipitation,separating the proteins with length 17 cm and pH 3-10 NL IPG strips,loading proteins samples of 150 μg followed by isoelectric focusing programⅠ,and selecting SDS-PAGE gel concentration at 12%.The 2-D images with good repeatability and high protein spot resolution were obtained.
作者 曾静 陈松 朱利泉 高启国 王小佳 杨晓红
机构地区 西南大学农学与生物技术学院 西南大学园艺园林学院
出处 《中国蔬菜》 北大核心 2012年第06X期30-36,共7页 China Vegetables
基金 国家自然科学基金项目(30900986 31071802) 重庆市自然科学基金项目(2009BB1298) 西南大学博士基金项目(SWUB2008042) 中央高校基本科研业务费专项(XDJK2010B010)
关键词 甘蓝柱头 自交不亲和 蛋白质双向电泳 Brassica oleracea L.var.capitata L.stigma Self-incompatibility Protein two-dimensional electrophoresis
分类号 S635 [农业科学—蔬菜学]
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参考文献12

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二级参考文献25

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共引文献3

同被引文献27

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二级引证文献7

中国蔬菜
2012年 第06X期

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