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.
出处
《园艺学报》
CAS
CSCD
北大核心
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)