大麦核仁蛋白基因HvNOP58的克隆和表达分析  

Cloning and Expression Characteristics of a Nucleolar Protein Gene HvNOP58 in Barley

作　　者：王梦玥 罗振蒙 詹开旺 毛孝强[1] 周洪斌 魏红霞 高洪铭 陈升位[1] WANG Mengyue;LUO Zhenmeng;ZHAN Kaiwang;MAO Xiaoqiang;ZHOU Hongbin;WEI Hongxia;GAO Hongming;CHEN Shengwei(College of Agronomy and Biotechnology,Yunnan Agricultural University,Kunming,Yunnan 650201,China;Plant Protection and Inspection Station of Zhanyi District,Qujing,Yunnan 655331,China)

机构地区：[1]云南农业大学农学与生物技术学院,云南昆明650201 [2]沾益区植保植检站,云南曲靖655331

出　　处：《麦类作物学报》2024年第9期1087-1095,共9页Journal of Triticeae Crops

基　　金：国家自然科学基金项目(32060457,32360469);云南省科技厅基础研究专项重点项目(202201AS070066)。

摘　　要：Nop(nucleolar protein)结构域蛋白可参与核糖体装配、核苷酸甲基化修饰、基因表达调控等生物进程,是制约真核生物发育遗传、细胞周期调控和衰老机制等研究的关键因素,但少有大麦Nop结构域核仁蛋白基因的报道。本研究以北青7号和Ynbs突变体为材料,克隆了大麦HvNOP58基因的CDS序列。结果表明,HvNOP58基因位于大麦Chr2H基因组负链的49019790~49023604 bp区间,具有10个外显子,编码区长度为1674 bp。所编码蛋白属于亲水性的稳定蛋白,其高级结构以α-螺旋和无规则卷曲为主,具有N端结构域、NOSIC结构域和Nop结构域。同源比对和系统进化分析表明,HvNOP58蛋白与同族物种的核仁蛋白具有更近的亲缘关系。亚细胞定位、RT-PCR检测和荧光定量检测结果表明,HvNOP58基因定位于细胞核,在叶、茎、幼穗、根和叶鞘5种组织中有表达。The protein with nucleolar protein(Nop)domain can participate in biological processes,such as ribosome assembly,nucleotide methylation modification and gene expression regulation,and is a key factor that restricts the development and inheritance,cell cycle regulation and aging mechanisms of eukaryotes.However,there were few reports on the nucleolar protein genes with Nop domain in barley.In the present study,the coding sequence of barley gene HvNOP58 was cloned using Beiqing 7 and Ynbs mutant as materials.The results showed that HvNOP58 gene was located within 49019790-49023604 bp range of the negative strand of the Chr2H in barley,and contained 10 exons,a coding region containing 1674 bp.The putative protein encoded by HvNOP58 belonged to a hydrophilic stable protein,and its advanced structure is characterized by someα-helices and random coils,containing the N-terminal domain,NOSIC domain and Nop domain.Homology comparison and phylogenetic analysis indicated that the HvNOP58 protein showed a closer phylogenetic relationship with nucleolar proteins of the same species.Based on subcellular localization,RT-PCR detection and fluorescence quantification,the HvNOP58 gene was located in the nucleolus and expressed of in the leaves,stems,young spikes roots,and leaf sheaths.

关 键 词：大麦 核仁蛋白 基因克隆 亚细胞定位 表达分析 

分 类 号：S512.3[农业科学—作物学] S330