毛果杨HDA902基因的克隆、序列分析和亚细胞定位  

Cloning, Sequence Analysis and Subcellular Localization of HDA902 Gene from Populus trichocarpa

作　　者：刘超[1] 李开隆[1] 刘春娟[1] 马旭俊[1] 

机构地区：[1]东北林业大学林木遗传育种国家重点实验室,哈尔滨150040

出　　处：《分子植物育种》2016年第5期1082-1090,共9页Molecular Plant Breeding

基　　金：黑龙江省博士后基金项目(LBH-Z13006);国家自然科学基金青年科学基金项目(31200497)共同资助

摘　　要：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） is an important enzyme which catalyzes histone deacetylation, regulates status of histone acetylation, and plays an important role in the modification of chromatin structure and the regulation of gene transcription. To study the subcellular localization of HDA 902 gene and lay the foundation for its further functional study, we cloned the encoded sequence of HDA 902 gene by PCR method, analyzed the encoded protein sequences by bioinformatics method, constructed the HDA902-GFP fusion gene expression vector and expressed it in the onion epidermal cells to determine the subcellular localization of HDA902. The results showed that the ORF of HDA902 gene was 1 500 bp, encoding a protein containing 499 amino acids. Sequence alignment and conserved domain analyses showed that HDA902 protein shared a conserved domain （HDAC3） with the HDACs in other plants. Phylogenetic analysis showed that HDA902 protein was relatively close to HDAC （XP_007200978.1） from Prunus persica and HDA19-1ike （XP_008348955.1） from Malus domestica in evolution. Promoter sequence analysis showed that the promoter sequence of HDA 902 gene contained multiple stress-related and light-responsive cis-acting elements, including 5UTR Py-rich stretch, ARE, ACE and HSE. Subcellular localization analysis showed that HDA902 protein mainly localized in the cytoplasm. These results would provide theoretical basis for further research on the function of HDA 902 gene in Populus trichocarpa.

关 键 词：毛果杨 组蛋白去乙酰化酶 基因克隆 生物信息学 亚细胞定位 

分 类 号：S792.11[农业科学—林木遗传育种]