青稞耐低氮相关类甜蛋白基因HvTOND1克隆和亚细胞定位研究

Cloning and Subcellular Localization of Related Thaumatin-like Protein Gene HvTOND1 Tolerant to Low Nitrogen in Hulless Barley

为了探索青稞耐低氮相关类甜蛋白基因HvTOND1的基因和蛋白结构特点,为青稞耐低氮分子机制研究提供基础。以青稞‘昆仑14’叶片为材料,根据植物基因组数据库Gramene(http://www.gramene.org/)中的大麦HvTOND1(HORVU5Hr1G005300)基因和启动子区域序列设计引物,通过PCR获得青稞‘昆仑14’HvTOND1基因和启动子区域序列。采用生物信息学软件对HvTOND1基因启动子区域元件以及蛋白理化性质、跨膜结构、磷酸化位点、信号肽、二级结构、三级结构进行分析。结果表明HvTOND1没有内含子,HvTOND1蛋白由171个氨基酸组成,具有4个磷酸化位点,具有信号肽,不具有跨膜结构。将HvTOND1与其他植物的氨基酸序列进行对比并构建进化树发现青稞HvTOND1与节节麦AtsTOND1亲缘关系最近。亚细胞定位结果显示HvTOND1定位在内质网中。

The gene and protein structure characteristics of HvTOND1 were explored for provideing a basis to understand the molecular mechanism of tolerance to low nitrogen in hulless barley.In this study,the leaves of hulless barley‘Kunlun 14’were used as materials,and primers were designed according to barley HvTOND1(HORVU5Hr1G005300)gene and promoter region sequences in Gramene(http://www.gramene.org/),and PCR was used to obtain the HvTOND1 gene and promoter region sequences of hulless barley‘Kunlun 14’.The bioinformatics software was used to analyze the HvTOND1 gene promoter region element and protein physical and chemical properties,transmembrane structure,phosphorylation sites,signal peptide,secondary structure and tertiary structure.The results showed that HvTOND1 had no intron,and HvTOND1 protein was composed of 171 amino acids,which had 4 phosphorylation sites,1 signal peptides and no transmembrane structure.By comparing HvTOND1 amino acid sequences with those of other plants,the phylogenetic tree was constructed.It is found thant the hulless barley HvTOND1 is closely related to AtsTOND1 and HvTOND1 is localized in the endoplasmic reticulum.