[1]庄维兵,高志红*,章镇,等.果梅花芽蛋白质双向电泳优化体系的建立[J].南京农业大学学报,2011,34(6):47.[doi:10.7685/j.issn.1000-2030.2011.06.009]
 ZHUANG Wei-bing,GAO Zhi-hong*,ZHANG Zhen,et al.Optimization of two-dimensional electrophoresis conditions for floral buds proteome analysis of Japanese apricot[J].Journal of Nanjing Agricultural University,2011,34(6):47.[doi:10.7685/j.issn.1000-2030.2011.06.009]
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果梅花芽蛋白质双向电泳优化体系的建立()
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《南京农业大学学报》[ISSN:1000-2030/CN:32-1148/S]

卷:
34卷
期数:
2011年6期
页码:
47
栏目:
出版日期:
2011-11-30

文章信息/Info

Title:
Optimization of two-dimensional electrophoresis conditions for floral buds proteome analysis of Japanese apricot
作者:
庄维兵高志红*章镇侍婷邵静
南京农业大学园艺学院,江苏 南京 210095
Author(s):
ZHUANG Wei-bingGAO Zhi-hong*ZHANG ZhenSHI TingSHAO Jing
College of Horticulture,Nanjing Agricultural University,Nanjing 210095,China
关键词:
果梅花芽蛋白提取双向电泳
Keywords:
Japanese apricotfloral budsprotein extractiontwo-dimensional electrophoresis(2-DE)
分类号:
S662.4
DOI:
10.7685/j.issn.1000-2030.2011.06.009
摘要:
以果梅花芽为材料,比较了TCA/丙酮法和酚抽法2种总蛋白质提取方法对蛋白质双向电泳(2-DE)的影响,同时对IPG胶条种类、凝胶浓度、蛋白质上样量及染色方法等环节进行了优化。结果表明:TCA/丙酮法比酚抽法在2-DE凝胶图谱上的蛋白质点数增多,图谱背景也比较清晰,没有明显的纵横条纹;采用17 cm pH 4~7的胶条、10%的凝胶浓度、1.5~1.8 mg的蛋白质上样量,经改良胶条考马斯亮蓝G-250染色后可获得背景清晰、重复性好、蛋白质分辨率高的2-DE图谱。高质量的果梅花芽蛋白质提取方法和蛋白质双向电泳体系的优化可为果梅蛋白质组学研究奠定技术基础。
Abstract:
The effects of different protein extraction methods on 2-DE maps were compared and the loading quantity of sample,the gel concentration and the staining methods were also optimized for the analysis of 2-DE system in Japanese apricot.The results showed that the samples prepared by TCA/acetone precipitation had been observed with more protein spots in the 2-DE maps than those prepared by phenol extraction method,clearer background and slighter streaks.And a high quality 2-DE map with low background was obtained using the following optimized procedure:loading 1.5-1.8 mg protein sample on 17 cm IPG strip with pH 4-7,SDS-PAGE with 10% gel concentration,and finally detecting proteins with modified colloidal coomassie brilliant blue G-250 staining.The optimized system would facilitate the proteomics research in Japanese apricot.

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备注/Memo

备注/Memo:
收稿日期:2011-01-26基金项目:农业部公益性行业(农业)科研专项经费(201003058);江苏省自然科学基金项目(BK2011642)作者简介:庄维兵,硕士研究生。*通讯作者:高志红,副教授,硕导,主要从事果树种质资源和分子生物学研究,E-mail:gaozhihong@njau.edu.cn。
更新日期/Last Update: 2011-12-06