大豆GmNF-YC4基因的克隆与亚细胞定位分析

doi:10.6048/j.issn.1001-4330.2025.06.008

新疆农业科学 ›› 2025, Vol. 62 ›› Issue (6): 1365-1370.DOI: 10.6048/j.issn.1001-4330.2025.06.008

• 作物遗传育种·耕作栽培·生理生化 • 上一篇下一篇

大豆GmNF-YC4基因的克隆与亚细胞定位分析

刘雷蕾¹(), 赵国勤¹, 聂俊芳², 于月华², 倪志勇¹()

1.新疆极端环境生物生态适应与进化重点实验室/新疆农业大学生命科学学院,乌鲁木齐, 830052
2.新疆农业大学农学院,乌鲁木齐 830052

收稿日期:2024-11-15 出版日期:2025-06-20 发布日期:2025-07-29
通信作者: 倪志勇(1981-),男,黑龙江人,教授,博士生导师,研究方向植物对非生物胁迫应答的分子机制,(E-mail)nizhiyong@126.com
作者简介:刘雷蕾(1998-),女,四川人,硕士研究生,研究方向生物学,(E-mail)1908181694@qq.com
基金资助:
国家自然科学基金项目(32160446);国家自然科学基金项目(32360454);“天山英才”培养计划(2023TSYCCX0012);新疆维吾尔自治区自然科学基金(2022D01E14);新疆维吾尔自治区级大学生创新创业训练计划项目(S202210758033)

Cloning and subcellular localization analysis of the GmNF-YC4 gene in soybean

LIU Leilei¹(), ZHAO Guoqin¹, NIE Junfang², YU Yuehua², NI Zhiyong¹()

1. Key Laboratory of Ecological Adaptation and Evolution of Extreme Environment in Xinjiang, College of Life Sciences, Xinjiang Agricultural University, Urumqi 830052, China
2. College of Agronomy, Xinjiang Agricultural University, Urumqi 830052, China

Received:2024-11-15 Published:2025-06-20 Online:2025-07-29
Supported by:
National Natural Science Foundation of China(32160446);National Natural Science Foundation of China(32360454);“Tianshan Talents”Training Program Project(2023TSYCCX0012);Natural Science Foundation of Xinjiang Uygur Autonomous Region(2022D01E14);College Students' Innovation and Entrepreneurship Training Program Project of Xinjiang Uygur Autonomous Region(S202210758033)

摘要/Abstract

摘要：

【目的】为了研究GmNF-YC4的亚细胞定位,为揭示大豆抗逆分子机理奠定基础。【方法】本研究从大豆中克隆基因GmNF-YC4,利用生物信息学对GmNF-YC4的序列进行分析并构建系统发生树,采用瞬时转化烟草方法分析GmNF-YC4在烟草细胞中的亚细胞定位。【结果】GmNF-YC4 (GenBank 登录号:NM_001362631.1)的开放阅读框大小为690 bp,编码229个氨基酸,预测分子量为24.78 kDa,等电点为5.19。GmNF-YC4蛋白含有一个HAP5保守结构域。GmNF-YC4和拟南芥的AtNF-YC1,AtNF-YC4聚在一个分枝中。GmNF-YC4的基因组包含1个外显子。GmNF-YC4蛋白定位于烟草的细胞核中。【结论】 GmNF-YC4是一个细胞核定位的转录因子,获得了GmNF-YC4基因序列。

关键词: 大豆; 核因子亚基C; 基因克隆; 亚细胞定位

Abstract:

【Objective】 To study the subcellular localization of GmNF-YC4,which lays the foundation for revealing the molecular mechanism of soybean stress resistance. 【Methods】 In this study, GmNF-YC4 was cloned from soybean, bioinformatics for GmNF-YC4 sequence and phylogenetic tree, and the subcellular localization of GmNF-YC4 in tobacco cells was analyzed by transient transformation tobacco. 【Results】 The open reading frame size of GmNF-YC4 (GenBank accession number: NM _ 001362631.1) was 690bp, encoding 229 amino acids, which predicted molecular weight of 24.78 kDa, and isoelectric point of 5.19. The GmNF-YC4 protein contained a HAP 5 conserved domain. The phylogenetic tree analysis showed that GmNF-YC4 and AtNF-YC 1, AtNF-YC 4 were clustered in one clade. The genome of GmNF-YC4 contained 1 exon. The subcellular localization results indicated that GmNF-YC4 protein was localized in the nucleus of tobacco. 【Conclusion】 GmNF-YC4 is a nucleus-localized transcription factor. GmNF-YC4 gene sequence is obtained.

Key words: soybean; nuclear factor subunit C; gene cloning; subcellular localization

中图分类号:

S565.1

刘雷蕾, 赵国勤, 聂俊芳, 于月华, 倪志勇. 大豆GmNF-YC4基因的克隆与亚细胞定位分析[J]. 新疆农业科学, 2025, 62(6): 1365-1370.

LIU Leilei, ZHAO Guoqin, NIE Junfang, YU Yuehua, NI Zhiyong. Cloning and subcellular localization analysis of the GmNF-YC4 gene in soybean[J]. Xinjiang Agricultural Sciences, 2025, 62(6): 1365-1370.

图/表 3

图1 GmNF-YC4序列注: a: GmNF-YC4 与拟南芥 NF-YC家族系统进化树分析; b: GmNF-YC4蛋白保守结构域预测及与拟南芥NF-YB家族氨基酸序列比对分析; c: GmNF-YC4蛋白保守结构域预测;d:GmNF-YC4结构示意图

Fig.1 Sequence of GmNF-YC4 Notes: a: analysis of the phylogenetic tree of GmNF-YC4 and the Arabidopsis NF-YC family; b: prediction of conserved domain of GmNF-YC4 protein and analysis of amino acid sequence alignment with the Arabidopsis NF-YB family; c: prediction of conserved domain of GmNF-YC4 protein; d: schematic diagram of GmNF-YC4 structure

图2 大豆GmNF-YC4基因克隆注: M: DL2000 DNA Marker (a).目的片段扩增;1:GmNF-YC4编码区扩增产物;(b).菌液PCR检测GmNF-YC4-pEGAD

Fig.2 Cloning of GmNF-YC4 gene Notes: M: DL2000 DNA Marker (a). Amplification of the target fragment; 1: the amplified product of the GmNF-YC4 coding region; (b). GmNF-YC4-pEGAD was detected by PCR of the bacterial solutio

图3 大豆GmNF-YC4亚细胞定位

Fig.3 GmNF-YC04 subcellular localization

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大豆GmNF-YC4基因的克隆与亚细胞定位分析

Cloning and subcellular localization analysis of the GmNF-YC4 gene in soybean

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