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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。
PAP3 (Gen Bank Accession No: HM104635) and PPI (Gen Bank Accession No: GU812176) are the B-type genes that control floral organ development in pepper. They control the development of petals and stamens and belong to the MADS-box family of genes. To verify the interaction between PAP3 protein and PPI protein, the yeast two-hybrid system was used to construct the yeast expression vectors pGBKT7-PPI and pGADT7-PAP3. After self-activation and toxicity were not transformed, the fusion diploid Yeast (pGBKT7-PPI × pGADT7-PAP3) was able to grow on auxotrophic medium, and the in vitro GST-Pull down technique was used to incubate the induced GST-PAP3 protein and His-PPI protein. The results showed that both PAP3 protein and There are specific interactions between PPI proteins.