水稻组蛋白去甲基化酶基因OsJMJ719的克隆及表达分析

Analysis on Cloning and Expression of Histone Demethylase Gene OsJMJ719 in Rice

【目的】克隆水稻组蛋白去甲基化酶OsJMJ719基因,分析其在非生物胁迫下的表达模式,为探究OsJMJ719在非生物胁迫响应中的功能提供理论依据。【方法】以水稻品种中花11为材料,克隆获得完整的OsJMJ719基因,对其进行生物信息学分析,构建OsJMJ719与绿色荧光蛋白(GFP)融合表达载体35S::OsJMJ719-GFP,利用烟草瞬时转化体系和水稻原生质体转化的方法来观察蛋白的亚细胞定位,并应用实时荧光定量PCR技术分析OsJMJ719基因在水稻不同组织中的表达情况以及在非生物胁迫处理下的表达模式。【结果】OsJMJ719基因(LOC_Os02g01940)编码区长度为2994 bp,编码997个氨基酸,OsJMJ719启动子区域含有10个植物激素响应元件和3个环境胁迫调控相关元件。系统进化分析结果显示,OsJMJ719与沼生菰、粗山羊草、小麦和大麦中的JMJ蛋白有较高的同源性。亚细胞定位结果显示,OsJMJ719蛋白定位于细胞核内。荧光定量结果显示,OsJMJ719在种子中表达量较高,且该基因的表达受ABA、NaCl和PEG6000的诱导,推测其在非生物胁迫过程中起重要作用。【结论】本研究展示了OsJMJ719基因的表达蛋白在进化树中的位置及其同源性物种,揭示了其表达蛋白定位于细胞中的具体位置、蛋白的结构及特征,以及该基因主要受到哪些外界因素调控,为进一步研究OsJMJ719基因的功能提供基础资料。

【Objective】A novel rice histone demethylase gene OsJMJ719 was cloned.The expression pattern of this gene under abiotic stress was analyzed to provide theoretical basis for exploring the function of OsJMJ719 in response to abiotic stress.【Method】The complete OsJMJ719 gene was obtained by cloning with ZH11 as material.35S::OsJMJ719-GFP expression vector integrated with OsJMJ719 and green fluorescent protein was established for subcellular localization.Transient tobacco expression system and rice protoplast transformation method was employed to observe the subcellular localization and real-time PCR was used to analyze the expression profile of OsJMJ719 in different tissues of rice and the expression pattern under abiotic stress treatments.【Result】The coding region of OsJMJ719 gene(LOC_Os02g01940)is 2994 bp in length,encoding 997 amino acids.The promoter region of OsJMJ719 contains 10 phytohormone response elements and 3 environmental stress regulation related elements.Phylogenetic analysis shows that OsJMJ719 has high homology with JMJ protein in wild Zizania palustris,Aegilops tauschii,Triticum aestivum and Hordeum vulgare.The subcellular localization results reveal that OsJMJ719 protein is localized in the nucleus.Fluorescence quantitative results reveal that OsJMJ719 is highly expressed in seeds,and the expression of this gene is induced by ABA,NaCl and PEG6000,suggesting that it plays an important role in the process of abiotic stress.【Conclusion】The study shows the location of OsJMJ719 in the phylogenetic tree and its homologous species,reveals the localization,structure and characteristics of the protein,and the main external factors responsible for the regulation of this gene.These results provide a fundamental data basis for further research on the function of OsJMJ719 gene.