| 利用酵母双杂交法检测甘蓝SCR与SRK之间的相互作用 |
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| 引用本文： | 罗兵,薛丽琰,朱利泉,张贺翠,彭一波,陈松,杨红,杨昆,李成琼,王小佳.利用酵母双杂交法检测甘蓝SCR与SRK之间的相互作用[J].作物学报,2011,37(4):579-586. |
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| 作者姓名： | 罗兵 薛丽琰 朱利泉 张贺翠 彭一波 陈松 杨红 杨昆 李成琼 王小佳 |
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| 作者单位： | 1 西南大学植物生理生物化学实验室, 重庆400716; 2 西南大学重庆市蔬菜学重点实验室, 重庆400716；3常熟理工学院生物与食品工程学院，常熟 215500 |
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| 摘 要： | 为深入研究甘蓝S位点受体激酶(SRK)和S位点富含半胱氨酸蛋白(SCR)间相互识别的分子机理，以具有典型自交不亲和性的甘蓝D3为材料，利用巢式PCR分别扩增SRK胞外域(eSRK)和SCR，通过酵母双杂交检测二者之间的相互作用。并利用同源重组技术将eSRK与GAL4报告基因DNA活化域融合(pGADT7eSRK)以及SCR与GAL4报告基因DNA结合域融合(pGBKT7SCR)。两种重组质粒分别转化酵母Y187和Y2HGold后未出现自激活现象。融合的二倍体酵母(pGADT7eSRK× pGBKT7SCR)能在选择性固体培养基(SD/–Ade/–His/–Leu/–Trp/X-a-Gal/AbA)上生长，并且菌落呈蓝色。结果表明, eSRK与SCR蛋白能够相互结合, 为深入研究二者作用位点成功建立了一个技术平台。
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| 关 键 词： | 甘蓝 S位点受体激酶(SRK) S位点富含半胱氨酸蛋白(SCR) 酵母双杂交 |
| 收稿时间： | 2010-09-17 |
Detection of Interactions between SCR and SRK in Brassica oleracea L.by Yeast Two-Hybrid System |
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| LUO Bing,XUE Li-Yan,ZHU Li-Quan,ZHANG He-Cui,PENG Yi-Bo,CHEN Song,YANG Hong,YANG Kun,LI Cheng-Qiong,WANG Xiao-Jia.Detection of Interactions between SCR and SRK in Brassica oleracea L.by Yeast Two-Hybrid System[J].Acta Agronomica Sinica,2011,37(4):579-586. |
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| Authors: | LUO Bing XUE Li-Yan ZHU Li-Quan ZHANG He-Cui PENG Yi-Bo CHEN Song YANG Hong YANG Kun LI Cheng-Qiong WANG Xiao-Jia |
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| Institution: | 1.Plant Physiology and Biochemistry Laboratory of Southwest University, Chongqing 400716, China;2.Key Laboratory in Olericulture of Chongqing, Southwest University, Chongqing 400716, China;3.College of Biology and Food Engineering, Changshu Institute of Technology, Changshu 215500, China |
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| Abstract: | For further study on mechanism of the mutual recognition between S-locus receptor kinase (SRK) and S-locus Cysteine-rich protein (SCR) in Brassica, the extracellular domains of SRK (named as eSRK) and SCR were amplified by nested PCR using Brassica oleracea L. D3 with the trait of typical self-incompatibility (SI), and the interactions between eSRK and SCR was detected by the yeast two-hybrid system. Then the full-length eSRK and the SCR were fused to the Gal4 DNA activation domain (designated pGADT7eSRK) a... |
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| Keywords: | Brassica oleracea L Self-incompatibility S-locus receptor kinase (SRK) S-locus cysteine-rich protein (SCR) Yeast two-hybrid system |
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