木薯MeSAUR1互作蛋白的筛选及鉴定

Screening and Identification of MeSAUR1 Interacting Protein in Cassava

木薯MeAGPS1a基因编码的小亚基是淀粉合成关键酶AGPase的催化中心,前期研究发现MeSAUR1转录因子可结合MeAGPS1a基因启动子并调控该基因表达。采用酵母双杂交技术筛选MeSAUR1转录因子的互作蛋白,可进一步解析MeAGPS1a基因表达的分子调控网络。本研究构建了MeSAUR1的酵母双杂交诱饵载体pGBKT7-MeSAUR1,采用酵母双杂交技术从木薯cDNA文库中筛选MeSAUR1的候选互作蛋白,结果共筛选出31个阳性克隆。通过酵母双杂交点对点实验验证了候选互作蛋白MePP2C与MeSAUR1的互作关系,本研究结果有助于揭示Me SAUR1蛋白调控木薯淀粉合成的互作网络。

The small subunit enco ded by MeAGPS1a gene is the catalytic center of AGPase in cassava,and AGPase is a key enzyme in starch synthesis.Previous studies have found that MeSAUR1 transcription factor can bind to the promoter of MeAGPS1a gene and regulate its expression.Screening the interacting protein of MeSAUR1 transcription factor by yeast two-hybrid technology can further analyze the molecular regulatory network of MeAGPS1a gene expression.The yeast two-hybrid vector pGBKT7-MeSAUR1 of MeSAUR1 was constructed in this study,and the candidate interacting proteins of MeSAUR1 were screened from cassava cDNA library by yeast two-hybrid technology,and 31 positive clones were screened.The interaction between candidate interacting protein MePP2C and MeSAUR1 was verified by yeast two-hybrid point-to-point experiment.The results of this study are beneficial to reveal the interacting network of MeSAUR1 protein regulating starch synthesis in cassava.