研究报告

酵母双杂交筛选与GsCBRLK相互作用的蛋白质

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  • 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

  网络出版日期: 2013-03-25

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

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  • Laboratory of Plant Bioengineering, College of Life Sciences, Northeast Agriculture University, Harbin 150030, China

Received date: 2012-09-11

  Revised date: 2013-01-25

  Online 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。

本文引用格式

杨姗姗,孙晓丽,于洋,才华,纪巍,柏锡,朱延明 . 酵母双杂交筛选与GsCBRLK相互作用的蛋白质[J]. 遗传, 2013 , 35(3) : 388 -394 . DOI: 10.3724/SP.J.1005.2013.00388

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.

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