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组蛋白去乙酰化酶(histone deacetylases,HDACs)是一类重要的酶,催化组蛋白去乙酰化,参与组蛋白乙酰化状态的调节,对染色体结构修饰和基因表达调控发挥着重要的作用。为了研究毛果杨HDA902基因的亚细胞定位,进而为研究其功能奠定基础,采用PCR的方法克隆了毛果杨组蛋白去乙酰化酶基因HDA902的编码序列,采用生物信息学方法分析了该基因所编码的蛋白质序列,并构建了HDA902-GFP融合基因表达载体,通过其在洋葱表皮细胞的表达来确定HDA902蛋白的亚细胞定位。研究结果表明,毛果杨HDA902基因的开放阅读框为1 500 bp,编码1个由499个氨基酸残基组成的蛋白质。序列比对和结构域分析显示,毛果杨HDA902蛋白与其他植物同源蛋白具有一个保守的结构域HDAC3。系统进化树分析表明,毛果杨HDA902蛋白在进化上与桃(Prunus persica)HDAC(XP_007200978.1)和苹果(Malus domestica)HDA19-like(XP_008348955.1)的亲缘关系较近。启动子分析表明,毛果杨HDA902基因的启动子序列包含5UTR Pyrich stretch、ARE、ACE和HSE等多个与逆境和光响应相关的顺式作用元件。亚细胞定位分析表明,毛果杨HDA902蛋白主要存在于细胞质中。这些研究结果为进一步研究毛果杨组蛋白去乙酰化酶HDA902基因的功能提供了理论基础。
Histone deacetylases (HDACs) are an important class of enzymes that catalyze histone deacetylation and are involved in the regulation of histone acetylation and play an important role in chromatin structure modification and gene expression regulation. In order to study the subcellular location of Populus trichocarpa on HDA902 gene and further study its function, the coding sequence of HDAC902 gene was cloned by PCR and the gene was analyzed by bioinformatics Encoded the protein sequence, and constructed HDA902-GFP fusion gene expression vector, through the expression of onion epidermal cells to determine the subcellular localization of HDA902 protein. The results show that Populus trichocarpa HDA902 gene open reading frame of 1500 bp, encoding a protein consisting of 499 amino acid residues. Sequence alignment and domain analysis showed that Populus trichocarpa HDA902 protein has a conserved domain HDAC3 with other plant homologous proteins. Phylogenetic tree analysis showed that the Populus tomentosa HDA902 protein is phylogenetically closer to Prunus persica HDAC (XP_007200978.1) and Malus domestica HDA19-like (XP_008348955.1). Promoter analysis showed that the promoter sequence of Populus tomentosa HDA902 gene contains many cis-acting elements related to stress and photoresponse, such as 5UTR Pyrich stretch, ARE, ACE and HSE. Subcellular localization analysis showed that Populus trichocarpa HDA902 protein mainly exists in the cytoplasm. These results provide a theoretical basis for further study on the function of the Populus tomentosa histone deacetylase HDA902 gene.