贾俊婷,陈兵先,吴柔贤,戴彰言,钟明生,解 昊,刘双幸,刘 军.水稻组蛋白去甲基化酶基因 OsJMJ719
的克隆及表达分析[J].广东农业科学,2022,49(11):152-161 |
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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 HistoneDemethylase Gene OsJMJ719 in Rice |
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DOI:10.16768/j.issn.1004-874X.2022.11.016 |
中文关键词: 水稻 OsJMJ719 组蛋白去甲基化 基因克隆 表达分析 |
英文关键词: rice OsJMJ719 histone demethylase gene cloning expression analysis |
基金项目:国家自然科学基金(31871716);广东省农业科学院院长基 金(202023);广州市科技计划项目
(202102021009);广东省农业科学院创新基金(202118) |
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中文摘要: |
【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 2 994 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。 |
英文摘要: |
【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
2 994 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. |
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