| 甘蓝柱头蛋白质双向电泳体系的建立 |
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| 引用本文： | 曾静,陈松,朱利泉,高启国,王小佳,杨晓红. 甘蓝柱头蛋白质双向电泳体系的建立[J]. 中国蔬菜, 2012, 1(12): 30-36 |
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| 作者姓名： | 曾静 陈松 朱利泉 高启国 王小佳 杨晓红 |
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| 作者单位： | 1.西南大学农学与生物技术学院，西南大学植物生理生物化学实验室，重庆 400716,2西南大学园艺园林学院，南方山地园艺学教育部重点实验室，重庆 400715 |
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| 基金项目： | 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) |
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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17 cm、pH 3～10 NL的IPG胶条，上样量150 μg，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。
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| 关 键 词： | 甘蓝柱头 自交不亲和 蛋白质双向电泳 |
Establishment of Two-dimensional Electrophoresis System of Brassica oleracea L.var.capitata L.Stigma Protein |
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| ZENG Jing,CHEN Song,ZHU Li-quan,GAO Qi-guo,WANG Xiao-jia,YANG Xiao-hong. Establishment of Two-dimensional Electrophoresis System of Brassica oleracea L.var.capitata L.Stigma Protein[J]. China Vegetables, 2012, 1(12): 30-36 |
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| Authors: | ZENG Jing CHEN Song ZHU Li-quan GAO Qi-guo WANG Xiao-jia YANG Xiao-hong |
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| Affiliation: | 1.College of Agronomy and Biotechnology，Southwest University，Plant Physiology and Biochemistry Laboratory，Southwest University，Chongqing 400716，China|2College of Horticulture and Landscape Architecture，Southwest University，Key Laboratory of Horticulture Science for Southern Mountainous Regions，Ministry of Education，Chongqing 400715，China |
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| Abstract: | 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. |
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| Keywords: | Brassica oleracea L.var.capitata L.stigma Self-incompatibility Protein two-dimensional electrophoresis |
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