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烟草硝酸盐转运蛋白NtNRT1.7基因的克隆、亚细胞定位及表达模式分析

Cloning, subcellular localization and expression pattern analysis of tobacco nitrate transporter NtNRT1.7 gene

  • 摘要: 为研究硝酸转运蛋白(Nitrate transporter, NRT)在烟草吸收和转运硝酸盐中的作用,以烟草K326的cDNA为模板,克隆了一个全长1 839 bp、编码612个氨基酸的硝酸盐转运蛋白基因,命名为NtNRT1.7。生物信息学分析表明烟草NtNRT1.7具有典型跨膜结构域。同源性分析结果显示,NtNRT1.7与拟南芥AtNRT1.7的同源性达57.23%。亚细胞定位结果表明该蛋白定位在细胞膜上,是一个典型的膜蛋白。启动子分析和不同胁迫处理的qRT-PCR结果显示,NtNRT1.7主要在成熟叶中高表达,并受低氮诱导抑制表达,受干旱、低温、高盐等胁迫诱导增强表达,表明NtNRT1.7基因可能参与非生物胁迫条件下硝酸盐的再利用。

     

    Abstract: In order to study the role of nitrate transporter (NRT) in the uptake and transport of nitrate in tobacco, a nitrate transporter gene encoding 612 amino acids with a total length of 1 839 bp was cloned using the cDNA of tobacco cultivar K326 as a template and named as NtNRT1.7. Bioinformatics analysis indicated that NtNRT1.7 had typical transmembrane domains. The homology analysis showed that the homology of NtNRT1.7 and Arabidopsis thaliana AtNRT1.7 was 57.23%. Subcellular localization results showed that this protein was located on the cell membrane and was a typical membrane protein. Promoter analysis and QRT-PCR results of different stress treatments indicated that NtNRT1.7 gene was highly expressed in mature leaves, its expression was suppressed by low nitrogen and enhanced by drought, low temperature, high salt and other stresses, which suggested that NtNRT1.7 gene might be involved in nitrate recycle under abiotic stresses.

     

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