玉米转脂蛋白基因ZmLTP3启动子互作蛋白的筛选与鉴定

Screening and identification of proteins interacting to the lipid transfer protein gene ZmLTP3 promoter from maize

转脂蛋白(Lipid transfer protein,LTP)广泛参与植物的各种生长发育进程及逆境响应,但对其表达调控规律的研究则鲜有报道。首先鉴定了ZmLTP3基因上游启动子的核心序列,进而构建了玉米(Zea mays L.)自交系B73的cDNA表达文库,克隆了ZmLTP3基因上游启动子区域,利用酵母单杂交技术筛选,筛选ZmLTP3基因启动子序列互作蛋白。通过测序和生物信息学分析,共筛选到13个互作蛋白cDNA。其中ZmLSD1具有典型的转录因子特征,属于C4类型锌指蛋白家族成员。EMSA检测结果表明,只有ZmLSD1蛋白可以与来自ZmLTP3基因启动子的DNA探针序列结合。上述结果表明ZmLSD1能与ZmLTP3基因启动子结合,并调控其表达的转录因子。

Lipid transfer protein(LTP)is widely involved in plant growth and development process and stress responses,but the regulation of LTPs expression has rarely been reported.First,the core sequence of ZmLTP3 gene upstream promoter were identified,then,the cDNA expression library of maize inbred line B73 was constructed,and the upstream promoter region of ZmLTP3 gene was cloned,and the ZmLTP3 gene sequence interaction proteins were screened by yeast one hybridization technology.A total of 13 interacting protein cDNAs were screened by sequencing and bioinformatics analysis.ZmLSD1 is a member of C4 zinc finger protein family with typical transcription factor characteristics.EMSA analysis show that only the ZmLSD1 protein can bind to the DNA probe sequence from the promoter of ZmLTP3 gene.These results indicate that ZmLSD1 is a transcription factor that can bind to the promoter of ZmLTP3 gene and regulate its expression.