遗传 ›› 2013, Vol. 35 ›› Issue (3): 388-394.doi: 10.3724/SP.J.1005.2013.00388

• 研究报告 • 上一篇    

酵母双杂交筛选与GsCBRLK相互作用的蛋白质

杨姗姗, 孙晓丽, 于洋, 才华, 纪巍, 柏锡, 朱延明   

  1. 东北农业大学生命科学学院 植物生物工程研究室, 哈尔滨 150030
  • 收稿日期:2012-09-11 修回日期:2013-01-25 出版日期:2013-03-20 发布日期:2013-03-25
  • 通讯作者: 朱延明 E-mail:ymzhu2001@yahoo.com.cn
  • 基金资助:

    国家自然科学基金项目(编号:31171578)资助

Identification of calcium/calmodulin-binding receptor-like kinase GsCBRLK-interactive proteins using yeast two-hybrid system

YANG Shan-Shan, SUN Xiao-Li, YU Yang, CAI Hu, JI Wei, BAI Xi, ZHU Yan-Ming   

  1. Laboratory of Plant Bioengineering, College of Life Sciences, Northeast Agriculture University, Harbin 150030, China
  • Received:2012-09-11 Revised:2013-01-25 Online:2013-03-20 Published:2013-03-25

摘要: GsCBRLK(calcium/calmodulin-binding receptor-like kinase from Glycine soja)在ABA及盐胁迫诱导的钙离子信号通路中起到关键的调节作用。为深入研究GsCBRLK蛋白的作用机制, 文章采用膜酵母双杂交系统, 以GsCBRLK为诱饵蛋白, 筛选与其相互作用的蛋白质。通过构建野生大豆盐胁迫条件下的cDNA文库、膜酵母双杂交系统筛选、复筛、回转验证、生物信息学分析以及酵母体内互作验证等手段, 最终获得2个(SNARE 和 14-3-3 蛋白)与GsCBRLK诱饵蛋白相互作用的蛋白质。

关键词: GsCBRLK, 酵母双杂交, 蛋白质相互作用, 生物信息学分析

Abstract: GsCBRLK (calcium/calmodulin-binding receptor-like kinase from Glycine soja) links ABA (abscisic acid)- and salt-induced calcium/calmodulin signal in plant cells. In order to study the molecular mechanismes of GsCBLRK, the salt-treated Glycine soja cDNA library was screened with pBT3-STE-CBRLK as bait plasmid using yeast two hybrid system. Two positive clones (SNARE and 14-3-3 protein) were identified by constructing cDNA library of wild soybean under salt treatment, membrane system yeast two hybrid screening, multiple screen, rotary validation, bioinformatic analysis and in-teraction identification in yeast.

Key words: GsCBRLK, yeast two-hybrid system, protein interaction, bioinformatic analysis

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