| 辣椒PAP3和PPI蛋白相互作用的鉴定 |
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| 引用本文： | 马宁,刘辰,王茹芳,沈火林.辣椒PAP3和PPI蛋白相互作用的鉴定[J].园艺学报,2016,43(3):564-568. |
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| 作者姓名： | 马宁 刘辰 王茹芳 沈火林 |
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| 作者单位： | 中国农业大学园艺学院，设施蔬菜生长发育调控北京市重点实验室，北京 100193 |
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| 基金项目： | ‘十二五’国家科技计划项目(2012BAD02B02)，现代农业产业技术体系北京市果类蔬菜创新团队项目(BAIC01-2016)，国家自然科学基金项目(31071806) |
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| 摘 要： | PAP3(Gen Bank登录号:HM104635)和PPI(Gen Bank登录号:GU812176)基因是控制辣椒花器官发育的B类基因,共同控制花瓣和雄蕊的发育,均属于MADS-box家族基因。为验证PAP3蛋白和PPI蛋白的相互作用,利用酵母双杂交系统,构建pGBKT7-PPI和pGADT7-PAP3酵母表达载体,转化相应酵母AH109感受态细胞后未出现自激活和毒性现象,融合的二倍体酵母(pGBKT7-PPI×pGADT7-PAP3)能在营养缺陷型培养基上生长;同时利用体外GST-Pull down技术,将诱导的GST-PAP3蛋白和His-PPI蛋白孵育,结果均表明,PAP3蛋白和PPI蛋白之间存在特异性相互作用。
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| 关 键 词： | 辣椒 PAP3 PPI MADS-box家族 酵母双杂交 体外GST-Pull down |
The Identification of Interaction with PAP3 and PPI Protein in Pepper |
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| MA Ning,LIU Chen,WANG Ru-fang,SHEN Huo-lin.The Identification of Interaction with PAP3 and PPI Protein in Pepper[J].Acta Horticulturae Sinica,2016,43(3):564-568. |
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| Authors: | MA Ning LIU Chen WANG Ru-fang SHEN Huo-lin |
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| Institution: | 1. Department of Cardiovascular Medicine in South Building, Chinese PLA General Hospital, Beijing 100853, China;
2. Institute of Atherosclerosis, Key Laboratory of Atherosclerosis in Universities of Shandong, Taishan Medical University, Taian 271000, China |
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| Abstract: | In MADS-box family，PAP3 and PPI gene jointly control the growth of petals and stamen. Both of them are class B genes，which in charge of dominating floral organ development. In order to verify PAP3 protein and PPI protein interaction furtherly，yeast two-hybrid system was used in this study，we constructed yeast expression vectors pGBKT7-PPI and pGADT7-PAP3 transformed into corresponding AH109 yeast cells，and made sure that they did not appear the self-activation and toxicity. The yeast zygote diploids（pGBKT7-PPI × pGADT7-PAP3）exhibit on selective agar plates；Meanwhile，we used fusion GST-PAP3 protein and His-PPI protein perform the vitro GST-Pull down technology in this study. The two experimental results showed that specific interactions between PAP3 protein and PPI protein. |
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| Keywords: | pepper PAP3 PPI MADS-box family Y2H GST-Pull down |
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