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柑橘叶片和果皮总蛋白质提取及双向电泳体系的建立
引用本文:邓贵明,曾继吾,姜波,钟云,周碧容,易干军,彭新湘. 柑橘叶片和果皮总蛋白质提取及双向电泳体系的建立[J]. 热带亚热带植物学报, 2012, 20(1): 19-25
作者姓名:邓贵明  曾继吾  姜波  钟云  周碧容  易干军  彭新湘
作者单位:1. 广东省农业科学院果树研究所,广州 510640;华南农业大学生命科学学院,广州 540642
2. 广东省农业科学院果树研究所,广州,510640
3. 华南农业大学生命科学学院,广州,540642
基金项目:国家科技支撑计划项目(2008BAD92B05);国家公益性行业(农业)科研专项(nyhyzx07-023);广东省科技计划项目(2008A020100022; 2008A040102002; 2009B04060002; 2009B020201010)资助
摘    要:以春甜橘(Citrus reticulata Blanco‘Chuntianju’)果皮和叶片为材料,分别用Tris-HCl、尿素/硫脲(Thi/Urea)、三氯乙酸/丙酮(TCA)和酚(Phe)等4种方法提取柑橘总蛋白质,从蛋白质产量、单向SDS-PAGE和双向电泳等方面进行比较。结果表明,4种方法的分离效果存在较大差异,不论是以柑橘叶片还是果皮为材料,均以TCA法最好,且双向电泳图谱分辨率较好,蛋白点清晰、均匀、基本没有条纹,且蛋白点多。这说明TCA法不仅能很好地去除柑橘果皮、叶片中存在的大量干扰物质,而且还能得到稳定的蛋白点。

关 键 词:柑橘  蛋白质组  提取  双向电泳
收稿时间:2011-03-14
修稿时间:2011-05-20

Extraction of Total Proteins from Leaves and Peels of Citrus reticulata and Establishment of Two-dimensional Electrophoresis System
DENG Gui-ming,ZENG Ji-wu,JIANG Bo,ZHONG Yun,ZHOU Bi-rong,YI Gan-jun and PENG Xin-xiang. Extraction of Total Proteins from Leaves and Peels of Citrus reticulata and Establishment of Two-dimensional Electrophoresis System[J]. Journal of Tropical and Subtropical Botany, 2012, 20(1): 19-25
Authors:DENG Gui-ming  ZENG Ji-wu  JIANG Bo  ZHONG Yun  ZHOU Bi-rong  YI Gan-jun  PENG Xin-xiang
Affiliation:1. Fruit Research Institute, Guangdong Academy of Agricultural Science, China; 2. College of Life Science, South China Agricultural University, China;1. Fruit Research Institute, Guangdong Academy of Agricultural Science, China; 2. College of Life Science, South China Agricultural University, China;Fruit Research Institute, Guangdong Academy of Agricultural Science, China;Fruit Research Institute, Guangdong Academy of Agricultural Science, China;Fruit Research Institute, Guangdong Academy of Agricultural Science, China;Fruit Research Institute, Guangdong Academy of Agricultural Science, China;College of Life Science, South China Agricultural University, China
Abstract:The total proteins in peels and leaves of Citrus reticuiata Blanco ’ Chuntianju ’ were extracted by four methods,such as Tris-HCl,Thi/Urea,trichloroacetic acid/pacetone precipitation(TCA) and phenol(Phe) extraction methods,respectively,and their effects were compared by protein production,one-way SDS-PAGE and two-dimensional electrophoresis(2-DE).The results showed that TCA method was the best for both leaves and peels of C.reticuiata ’ Chuntianju’.The 2-DE map of proteins extracted by TCA method had high resolution,the protein spots were clear and uniform without stripes,and the number of protein spots were large.It suggested that the TCA method could not only remove the interfering substances in peels and leaves of C.reticuiata ’Chuntianju’,but also get more stable protein spots.
Keywords:Citrus reticuiata  Proteome  Exteaction  Two-dimensional electrophoresis (2-DE)
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