天津农学院学报 ›› 2019, Vol. 26 ›› Issue (4): 16-19.doi: 10.19640/j.cnki.jtau.2019.04.004

• 研究与简报 • 上一篇    下一篇

蓝莓休眠花芽蛋白质双向电泳体系的建立和优化

吕晨菲, 王茂思, 黄唯子, 杨静慧通信作者, 刘艳军, 黄俊轩   

  1. 天津农学院 园艺园林学院,天津300384
  • 收稿日期:2019-05-05 发布日期:2020-01-19
  • 通讯作者: 杨静慧(1961-),女,教授,博士,研究方向:园艺植物栽培、抗逆生理和分子育种研究。 E-mail:jinghuiyang2@aliyun.com。
  • 作者简介:吕晨菲(1995-),女,硕士在读,研究方向:果树引种、栽培研究。E-mail:1787410311@qq.com。
  • 基金资助:
    天津市科学技术局项目(16YFZCNC00750);天津市重大农业技术推广项目(2017CK0184);天津市林果现代产业技术体系创新团队项目(ITTFPRS2018002);天津市科技特派员项目(17ZXBFNC0031,18JCTPJC68000)

Establishment and optimization of two-dimensional electrophoresis system for proteins in dormant flower-bud of blueberry

LÜ Chen-fei, WANG Mao-si, HUANG Wei-zi, YANG Jing-huiCorresponding Author, LIU Yan-jun, HUANG Jun-xuan   

  1. College of Horticulture and Landscape, Tianjin Agricultural University, Tianjin 300384, China
  • Received:2019-05-05 Published:2020-01-19

摘要: 为建立适合蓝莓花芽的蛋白质双向电泳技术体系,以天津市蓟州区主栽蓝莓品种‘布里吉塔’休眠花芽为试材,采用TCA/丙酮法提取蓝莓休眠花芽蛋白质,选择5种蛋白质上样量(2.00、2.25、2.50、2.75、3.00 mg)、2种聚焦电压(8 000、10 000 V)、3种聚焦时间(6、7、8 h)进行双向电泳,R-250考染后,进行图谱比较分析。结果表明,蛋白质上样量3.00 mg,聚焦电压8 000 V,聚焦时间为8 h,可得到蛋白数目多、分辨率高的蓝莓休眠花芽双向电泳图谱。建立了适合蓝莓休眠花芽蛋白质的双向电泳体系,为开展蓝莓蛋白质组学分析研究奠定了基础,更为蓝莓花芽休眠及解除休眠分子机制的研究提供了理论依据与技术支撑。

关键词: 蓝莓, 花芽, 蛋白质, 双向电泳

Abstract: In order to establish a protein two-dimensional electrophoresis system suitable for blueberry flower -bud, taking the dormantflower bud of 'Brigita', the main blueberry variety in Jizhou District of Tianjin as the test material, selectingfive protein samples volumes(2.00, 2.25, 2.50, 2.75, 3.00 mg), two focusing voltages(8 000, 10 000 V)and three focusing times(6, 7, 8 h)for two-dimensional electrophoresis,the chromatographic patterns after R-250 Coomassie brilliant blue dyeingwere compared and analyzed.. The results showed that, when the sample volumes was 3.00 mg, the focusing voltage was 8000 V and the focusing time was 8 h, which can obtain a high resolution electrophoresis map with a large number of protein spots. A two- dimensional electrophoresis system suitable for blueberry dormant flower bud protein was established, which laysa foundation for the development of blueberry proteomics analysis, and providestheoretical basis and technical support for the study of molecular mechanism of blueberry flower bud dormancy and dormancy release.

Key words: blueberry, flower bud, protein, two-dimensional electrophoresis

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