| 酵母双杂交筛选与GsCBRLK相互作用的蛋白质 |
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| 引用本文： | 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. |
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| 作者姓名： | 杨姗姗 孙晓丽 于洋 才华 纪巍 柏锡 朱延明 |
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| 作者单位： | 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 |
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| 基金项目： | 国家自然科学基金项目(编号：31171578)资助 |
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| 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。
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| 关 键 词： | GsCBRLK 酵母双杂交 蛋白质相互作用 生物信息学分析 |
| 收稿时间： | 2012-09-11 |
Identification of calcium/calmodulin-binding receptor-like kinase GsCBRLK-interactive proteins using yeast two-hybrid system |
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| Institution: | Laboratory of Plant Bioengineering, College of Life Sciences, Northeast Agriculture University, Harbin 150030, China |
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| 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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| Keywords: | GsCBRLK yeast two-hybrid system protein interaction bioinformatic analysis |
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