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拟南芥转录因子NF-YC的原核表达及多克隆抗体制备 被引量:7
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作者 卫静 李长江 +2 位作者 刘亚静 黄桂雪 张凌云 《生物技术通报》 CAS CSCD 北大核心 2012年第3期57-62,共6页
采用PCR方法扩增NF-YC基因得到其全长cDNA序列,并将其克隆至原核表达载体pET-48b中,在大肠杆菌BL21中用IPTG诱导出分子量约为45 kD的融合蛋白,SDS-PAGE和Western blotting检测鉴定表达产物。利用亲和层析技术对融合蛋白进行纯化,纯化后... 采用PCR方法扩增NF-YC基因得到其全长cDNA序列,并将其克隆至原核表达载体pET-48b中,在大肠杆菌BL21中用IPTG诱导出分子量约为45 kD的融合蛋白,SDS-PAGE和Western blotting检测鉴定表达产物。利用亲和层析技术对融合蛋白进行纯化,纯化后的目的蛋白免疫新西兰兔制备多克隆抗体。间接ELISA检测抗体效价大于1 62 500,Western blotting结果显示,该抗体可特异性识别NF-YC蛋白。 展开更多
关键词 拟南芥 转录因子 nf-yc融合蛋白 原核表达 多克隆抗体
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大麦NF-YC基因鉴定及在盐胁迫下的表达分析 被引量:2
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作者 王菲菲 周振祥 +5 位作者 洪益 谷洋洋 吕超 郭宝健 朱娟 许如根 《植物学报》 CAS CSCD 北大核心 2023年第1期140-149,共10页
核因子Y(NF-Y)是由NF-YA、NF-YB和NF-YC三个亚基组成的一类真核细胞转录因子,主要参与植物生长发育调控和非生物胁迫信号传递。该研究利用生物信息学方法解析了大麦(Hordeum vulgare)NF-YC基因家族功能。首先,基于大麦基因组数据库鉴定... 核因子Y(NF-Y)是由NF-YA、NF-YB和NF-YC三个亚基组成的一类真核细胞转录因子,主要参与植物生长发育调控和非生物胁迫信号传递。该研究利用生物信息学方法解析了大麦(Hordeum vulgare)NF-YC基因家族功能。首先,基于大麦基因组数据库鉴定出11个HvNF-YC成员,分布在除第2号染色体以外的其余6条染色体上,内含子0–5个。系统进化分析显示,大麦、拟南芥(Arabidopsis thaliana)和水稻(Oryza sativa)NF-YC基因家族成员可分为5个亚家族。基因复制分析显示,6个HvNF-YC基因存在片段复制,3个HvNF-YC基因存在串联复制。启动子顺式作用元件分析显示,大多数HvNF-YC基因启动子含有与非生物胁迫及激素响应相关的顺式作用元件。对HvNF-YC家族成员在不同组织不同时期的表达模式分析表明,不同成员的时空表达存在明显差异,其中HvNF-YC9和HvNF-YC11可能在籽粒发育初期发挥重要作用。通过分析耐盐型和盐敏感型大麦品种根和叶中HvNF-YC表达量变化,发现HvNF-YC3、HvNF-YC6和HvNF-YC10主要在盐胁迫初期的根中行使功能,HvNF-YC9主要在长期盐胁迫处理后期的根中起作用。综上所述,推测HvNF-YC9、10、11三个基因可作为后续探究大麦NF-YC参与耐盐作用机制的候选基因。该研究结果为进一步解析HvNF-YC在大麦中的耐盐调控功能奠定了基础。 展开更多
关键词 nf-yc 基因家族分析 大麦 盐胁迫 基因表达
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小麦NF-Y型转录因子基因TaNF-YC1介导烟草植株抵御渗透协迫功能研究 被引量:1
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作者 马自飞 宋文腾 +3 位作者 史美华 白菁华 路文静 肖凯 《河北农业大学学报》 CAS CSCD 北大核心 2020年第5期8-14,共7页
渗透胁迫逆境限制植株生长和发育,增强作物渗透胁迫抵御功能有助于实现作物高产优质目标。本研究以应答渗透逆境小麦NF-YC家族转录因子基因TaNF-YC1为基础,旨在阐明该基因在介导烟草植株抵御干旱和盐分逆境中的功能。系统进化分析表明,T... 渗透胁迫逆境限制植株生长和发育,增强作物渗透胁迫抵御功能有助于实现作物高产优质目标。本研究以应答渗透逆境小麦NF-YC家族转录因子基因TaNF-YC1为基础,旨在阐明该基因在介导烟草植株抵御干旱和盐分逆境中的功能。系统进化分析表明,TaNF-YC1与部分植物种属NF-Y家族成员在核苷酸序列组成上高度相似。干旱和盐分胁迫处理下TaNF-YC1的转录水平均明显上调,且逆境诱导增多的该基因转录本在正常恢复处理下不断减少。通过检测农杆菌介导烟草叶片转化的融合蛋白信号特征,表明TaNF-YC1蛋白经内质网分选后定位于细胞核内。与野生型相比,干旱和盐分处理下,正义表达TaNF-YC1烟草转基因植株长势改善、干重增加、光合碳同化能力增强;与此相反,反义表达烟草转基因植株渗透胁迫处理下的植株长势变差、干重降低、光合能力减弱。干旱处理下,正、反义转基因植株叶片气孔关闭速率分别呈加快和缓慢特征。本研究发现,TaNF-YC1对渗透逆境产生明显应答,通过调控气孔关闭影响渗透胁迫下植株光合和物质生产能力,在介导植株抵御渗透胁迫逆境中发挥重要功能。 展开更多
关键词 小麦 nf-yc转录因子 渗透胁迫 基因表达 遗传转化 功能分析
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拟南芥NF-YC9重组蛋白的制备
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作者 张晓娅 张国芳 +2 位作者 司琪 崔素娟 赵红桃 《河北农业大学学报》 CAS CSCD 北大核心 2016年第2期15-20,共6页
NF-YC9是模式植物拟南芥核因子Y家族成员之一,在开花调控等过程中发挥重要作用,但其蛋白的特性及其作用的分子机制尚待探究。本文首先扩增获得了NF-YC9基因的序列,采用重组技术构建了与麦芽糖结合蛋白MBP编码基因融合的重组表达载体,将... NF-YC9是模式植物拟南芥核因子Y家族成员之一,在开花调控等过程中发挥重要作用,但其蛋白的特性及其作用的分子机制尚待探究。本文首先扩增获得了NF-YC9基因的序列,采用重组技术构建了与麦芽糖结合蛋白MBP编码基因融合的重组表达载体,将其转化到大肠杆菌BL21中,进行了MBP-NF-YC9融合蛋白的诱导表达,并借助直链淀粉亲和层析介质纯化技术获得了电泳纯的MBP-NF-YC9重组蛋白,以备进一步对NF-YC9蛋白的生化特性进行全面检测。 展开更多
关键词 拟南芥 nf-yc9 重组蛋白 工程菌 MBP 亲和纯化
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玉米NF-YC亚基基因的鉴定与表达规律分析
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作者 王雪 李瑾 +5 位作者 朱丽 张悦 藏金萍 张康 邢继红 董金皋 《植物病理学报》 CAS CSCD 北大核心 2022年第1期123-128,共6页
0引言NF-Y(nuclear factor-Y,NF-Y)是一类与CCAAT基序结合的转录因子,由NF-YA、NF-YB和NF-YC 3个亚基组成[1-2]。NF-YC参与并调控植物生长发育的进程以及在生物和非生物胁迫响应过程中起作用[3]。AtNF-YC1通过与AtXTH21基因启动子区的CC... 0引言NF-Y(nuclear factor-Y,NF-Y)是一类与CCAAT基序结合的转录因子,由NF-YA、NF-YB和NF-YC 3个亚基组成[1-2]。NF-YC参与并调控植物生长发育的进程以及在生物和非生物胁迫响应过程中起作用[3]。AtNF-YC1通过与AtXTH21基因启动子区的CCAAT-box结合调控拟南芥对冷胁迫的响应[4]。AtNF-YC4可响应病原菌的侵染,其过表达植株对病原菌的敏感性降低[5]。 展开更多
关键词 冷胁迫 生物和非生物胁迫 转录因子 表达规律 NF 亚基基因 响应过程
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Arabidopsis NF-YCs Mediate the Light-Controlled Hypocotyl Elongation via Modulating Histone Acetylation 被引量:16
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作者 Yang Tang Xuncheng Liu +3 位作者 Xu Liu Yuge Li Keqiang Wu Xingliang Hou 《Molecular Plant》 SCIE CAS CSCD 2017年第2期260-273,共14页
Light is a crucial environmental signal that promotes photomorphogenesis, the developmental process with a series of light-dependent alterations for plants to adapt various external challenges. Chromatin modification ... Light is a crucial environmental signal that promotes photomorphogenesis, the developmental process with a series of light-dependent alterations for plants to adapt various external challenges. Chromatin modification has been proposed to be involved in such light-mediated growth, but the underlying mecha- nism is still elusive. In this study, we identified four Arabidopsis thaliana Nuclear Factor-YC homologs, NF- YCl, NF-YC3, NF-YC4, and NF-YC9 (NF-YCs), which function redundantly as repressors of light-controlled hypocotyl elongation via histone deacetylation. Obvious etiolation phenotypes are observed in NF-YCs loss-of-function mutant seedlings grown under light conditions, including significant elongated hypocotyls and fewer opened cotyledons. We found that NF-YCs interact with histone deacetylase HDA15 in the light, co-target the promoters of a set of hypocotyl elongation-related genes, and modulate the levels of histone H4 acetylation on the associated chromatins, thus repressing gene expression. In contrast, NF-YC-HDA15 complex is dismissed from the target genes in the dark, resulting in increased level of H4 acetylation and consequent etiolated growth. Further analyses revealed that transcriptional repression activity of NF-YCs on the light-controlled hypocotyl elongation partially depends on the deacetylation activity of HDA15, and loss of HDA15 function could rescue the short-hypocotyl phenotype of NF-YCs overexpression plants. Taken together, our results indicate that NF-YCl, NF-YC3, NF-YC4, and NF-YC9 function as tran- scriptional co-repressors by interacting with HDA15 to inhibit hypocotyl elongation in photomorphogen- esis during the early seedling stage. Our findings highlight that NF-YCs can modulate plant development in response to environmental cues via epigenetic regulation. 展开更多
关键词 PHOTOMORPHOGENESIS hypocotyl elongation nf-yc histone deactylation
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Arabidopsis ABF3 and ABF4 Transcription Factors Act with the NF-YC Complex to Regulate SOC1 Expression and Mediate Drought-Accelerated Flowering 被引量:19
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作者 Keumbi Hwang Hendry Susila +2 位作者 Zeeshan Nasim Ji-Yul Jung Ji Hoon Ahn 《Molecular Plant》 SCIE CAS CSCD 2019年第4期489-505,共17页
The drought-escape response accelerates flowering in response to drought stress, allowing plants to adaptively shorten their life cycles. Abscisic acid (ABA) mediates plant responses to drought, but the role of ABA-re... The drought-escape response accelerates flowering in response to drought stress, allowing plants to adaptively shorten their life cycles. Abscisic acid (ABA) mediates plant responses to drought, but the role of ABA-responsive element (ABRE)-binding factors (ABFs) in the drought-escape response is poorly understood. Here, we show that Arabidopsis thaliana ABF3 and ABF4 regulate flowering in response to drought through transcriptional regulation of the floral integrator SUPPRESSOR OF OVEREXPRESSION OF CONSTANS1 (SOC1). The abf3 abf4 mutant displayed ABA-insensitive late flowering under long-day conditions. Ectopic expression of ABF3 or ABF4 in the vasculature, but not in the shoot apex, induced early flowering, whereas expression of ABF3 fused with the SRDX transcript!onal repressor domain delayed flowering. We identified SOC1 as a direct downstream target of ABF3/4, and found that SOC1 mRNA levels were lower in abf3 abf4 than in wild-type plants. Moreover, induction of SOC1 by ABA was hampered in abf3 abf4 mutants. ABF3 and ABF4 were enriched at the -1028- to -657-bp region of the SOC1 promoter, which does not contain canonical ABF-ABRE-binding motifs but has the NF-Y binding element. We found that ABF3 and ABF4 interact with nuclear factor Y subunit C (NF-YC) 3/4/9 in vitro and in planta, and induction of SOC1 by ABA was hampered in nf-yc3 yc4 yc9 mutants. Interestingly, the abf3 abf4, nf-yc3 yc4 yc9, and sod mutants displayed a reduced drought-escape response. Taken together, these results suggest that ABF3 and ABF4 act with NF-YCs to promote flowering by inducing SOC1 transcription under drought conditions . This mechanism might contribute to adaptation by enabling plants to complete their life cycles under drought stress. 展开更多
关键词 DROUGHT ESCAPE abscisic acid ABF3 ABF4 nf-yc SOC1
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NF-YCs modulate histone variant H2A.Z deposition to regulate photomorphogenic growth in Arabidopsis 被引量:4
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作者 Chunyu Zhang Qian Qian +3 位作者 Xiang Huang Wenbin Zhang Xu Liu Xingliang Hou 《Journal of Integrative Plant Biology》 SCIE CAS CSCD 2021年第6期1120-1132,共13页
Inplants,lightsignalstriggeraphotomorphogenic program involving transcriptome changes, epigenetic regulation, and inhibited hypocotyl elongation. The evolutionarily conserved histone variant H2 A.Z, which functions in... Inplants,lightsignalstriggeraphotomorphogenic program involving transcriptome changes, epigenetic regulation, and inhibited hypocotyl elongation. The evolutionarily conserved histone variant H2 A.Z, which functions in transcriptional regulation, is deposited in chromatin by the SWI2/SNF2-RELATED 1 complex(SWR1 c). However, the role of H2 A.Z in photomorphogenesis and its deposition mechanism remain unclear. Here, we show that in Arabidopsis thaliana, H2 A.Z deposition at its target loci is induced by light irradiation via NUCLEAR FACTOR-Y, subunit C(NF-YC) proteins, thereby inhibiting photomorphogenic growth. NF-YCs physically interact with ACTIN-RELATED PROTEIN6(ARP6), a key component of the SWR1 c that is essential for depositing H2 A.Z, in a lightdependent manner. NF-YCs and ARP6 function together as negative regulators of hypocotyl growth by depositing H2 A.Z at their target genes during photomorphogenesis. Our findings reveal an important role for the histone variant H2 A.Z in photomorphogenic growth and provide insights into a novel transcription regulatory node that mediates H2 A.Z deposition to control plant growth in response to changing light conditions. 展开更多
关键词 ARABIDOPSIS LIGHT hypocotyl elongation nf-yc ARP6 H2A.Z deposition
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