辣椒PAP3和PPI蛋白相互作用的鉴定  

The Identification of Interaction with PAP3 and PPI Protein in Pepper

作　　者：马宁[1] 刘辰[1] 王茹芳[1] 沈火林[1] 

机构地区：[1]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

出　　处：《园艺学报》2016年第3期564-570,共7页Acta Horticulturae Sinica

基　　金：‘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)

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。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.

关 键 词：辣椒 PAP3 PPI MADS-box家族 酵母双杂交 体外GST-Pull DOWN 

分 类 号：S641.3[农业科学—蔬菜学]