烟草硝酸还原酶基因NIA1启动子互作蛋白的筛选

Screening of interactive proteins of nitrate reductase gene NIA1 promoter in Nicotiana tabacum

硝酸还原酶是植物氮同化过程中的关键酶和限速酶。为更好地调控硝酸还原酶基因的表达,本实验克隆了烟草硝酸还原酶基因NIA1上游938bp的启动子区域,构建了烟草cDNA基因表达文库,应用酵母单杂交技术筛选NIA1启动子序列互作蛋白。通过序列测定和生物信息学分析,共筛选出9个互作蛋白cDNA。其中,功能不明确的预测蛋白7个,具有明确功能信息的蛋白2个,分别为含有CRM结构域的CFM3蛋白(No. 71),和烟草亚硫酸盐氧化酶类似蛋白Sul?te oxidase-like蛋白(No. 126)。这些蛋白均为NIA1启动子序列互作蛋白,可能作为转录因子参与NIA1基因的转录调控过程。

Nitrate reductase is a key rate-limiting enzyme in nitrogen assimilation process in plants. In order to regulate gene expression of nitrate reductase, promoter of nitrate reductase gene NIA1 in tobacco which is 938 nucleotides in length was cloned and sequenced. In order to obtain information on interactive proteins which are probably interacted with the NIA1 promoter region, a tobacco cDNA gene expression library was constructed and yeast one hybrid technology was used to screen these proteins. Nine interactive proteins were identi?ed through sequencing and bioinformatics analysis. Among them 7 were identi?ed as uncharacterized proteins and 2 were annotated as functional proteins which are CFM3 protein(No.71) and sul?te oxidase-like protein(No.126). These identi?ed proteins are all NIA1 promoter interactive proteins which may act as transcriptional factors and be involved in the transcriptional regulation of NIA1 gene.