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Influence of High Temperature on the Expression of Arabidopsis Heat Shock Transcription Factor AtHsfA1a 被引量:1
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作者 Lihong GUO Juan CAI +1 位作者 Zhixuan YU Zhicuo DAN 《Agricultural Biotechnology》 CAS 2013年第1期13-15,共3页
[ Objective] This study ~med to investigate the influence of high temperature on the expression of heat shock transcription factor AtHsfAla in different genotypes of Arabidopsis. [ Method ] Arabidopsis plants overexpr... [ Objective] This study ~med to investigate the influence of high temperature on the expression of heat shock transcription factor AtHsfAla in different genotypes of Arabidopsis. [ Method ] Arabidopsis plants overexpressing heat shock transcription factor AtHsfA1 a were used as experimental materials and treated un- der high temperature at 39℃ for 1 rain and 5 min; total RNA of AtI-IsfAla was extracted, and the reverse transcription and amplification were conducted using RT- PCR technology, the amplification products were detected by electrophoresis. [ Result ] The expression levels of AtHsfA1 a in Arabidopsis plants overexpressing heat shock transcription factor AtHsfAla at high temperature and room temperature were higher than wild-type Arabidopsis; the expression levels of AtHsfAla in both wild-type Arab/dops/s and transgenic Arabidopsls plants overexpressing heat shock transcription factor AtHsfAla at high temperature of 39 ~C were higher than that at room temperature of 25 ~C, but the expression levels of AtHsfAla in wild-type Arab/dops/s and transgenic Arab/dops/s plants overexpressing heat shock transcription factor AtHsfAla varied little after high temperature treatment at 39 ~C for 1 rain or 5 rain. [ Conclusion] The expression of AtHsfAla is induced rapidly by high tem- perature, thus regulating the expression of early adversity-resistant genes. This study will lay the foundation demonstrating the mechanism of Arabidopsis heat shock transcription factor AtHsfAla. 展开更多
关键词 High temperature Arab/dops/s heat shock transcription factor AthsfAla Gene expression
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Prokaryotic Expression and Purification of Heat Shock Factor HSF1 in Arabidopsis thaliana
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作者 郭丽红 王定康 +3 位作者 袁燕 刘开庆 陈雪 陈善娜 《Agricultural Science & Technology》 CAS 2009年第4期41-42,145,共3页
[ Objective ] This study was to express and purify Arabidopsis thaliana heat shock factor HSF1. [ Method ] Using Escherichia coli M15 harboring HSF1 (pQE32/His6-HSF1, pREP4) as experimental materials, HSF1 was induc... [ Objective ] This study was to express and purify Arabidopsis thaliana heat shock factor HSF1. [ Method ] Using Escherichia coli M15 harboring HSF1 (pQE32/His6-HSF1, pREP4) as experimental materials, HSF1 was induced to express with isopropyl-β-D-galactoside (IPTG) ; then the expression product was purified using Ni-NTA-agarose affinity chromatography and analyzed by SDS-PAGE. [Result] HSF1 of Arabidopsis thaliana was successfully expressed and purified. [ Conclusion] This study provides materials for understanding the blinding site of HSF1 on Arabidopsis thaliana chromosome, further laying a good foundation for revealing the regulatory mechanism and physiological function of HSF1. 展开更多
关键词 heat shock factor hsf1 EXPRESSION PURIFICATION
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The ZmHSF08-ZmUGT92A1 module regulates heat tolerance by altering reactive oxygen species levels in maize 被引量:1
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作者 Hao Li Jing Wang +4 位作者 Menglong Li Leiming Wu Wenhui Rao Xiaojian Peng Haiyang Jiang 《The Crop Journal》 SCIE CSCD 2024年第5期1437-1446,共10页
GTs(Glycosyltransferases)are important in plant growth and abiotic stresses.However,its role in maize heat response is far from clear.Here,we describe the constitutively expressed UDP-glycosyltransferase ZmUGT92A1,whi... GTs(Glycosyltransferases)are important in plant growth and abiotic stresses.However,its role in maize heat response is far from clear.Here,we describe the constitutively expressed UDP-glycosyltransferase ZmUGT92A1,which has a highly conserved PSPG box and is localized in chloroplasts,is induced under heat stress.Functional disruption of ZmUGT92A1 leads to heat sensitivity and reactive oxygen species accumulation in maize.Metabolomics analysis revealed that ZmUGT92A1 affected multiple metabolic pathways and altered the metabolic homeostasis of flavonoids under heat stress.In vitro assay showed ZmUGT92A1 exhibits glycosyltransferase activity on flavonoids and hormones.Additionally,we identified a rapidly heat-induced transcription factor,ZmHSF08,which can directly bind and repress the promoter region of ZmUGT92A1.The ZmHSF08 overexpression line exhibits heat sensitivity and reactive oxygen species accumulation.These findings reveal that the ZmHSF08-ZmUGT92A1 module plays a role in heat tolerance in maize and provide candidate strategies for the development of heat-tolerant varieties. 展开更多
关键词 heat shock transcription factors heat stress Oxidative damage Uridine diphosphate glycosyltransferase Zea mays L.
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Advance in Research on Biological Function and TranscriptionalControl of Heat Shock Proteins 被引量:1
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作者 徐存拴 辛泽华 袁金云 《Developmental and Reproductive Biology》 2002年第2期130-136,共7页
The regulation of heat shock transcription factor to heat shock protein expression and the newest knowledge about the effect of heat shock protein on aging,immune response and the balance of cell survival and apoptosi... The regulation of heat shock transcription factor to heat shock protein expression and the newest knowledge about the effect of heat shock protein on aging,immune response and the balance of cell survival and apoptosis are summarized in the paper. 展开更多
关键词 heat shock protein (HSP) heat shock transcription factor (hsf) aging immune response the balance of cell survival and apoptosis
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Requirement for endogenous heat shock factor 1 in inducible nitric oxide synthase induction in murine microglia
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《中国药理学通报》 CAS CSCD 北大核心 2015年第B11期76-77,共2页
Aim Inducible nitric oxide synthase (iNOS) makes a great contribution to host defense and inflamma-tion. In many settings, lipopolysaccharide (LPS) induces iNOS expression through activation of the inhibitor of K... Aim Inducible nitric oxide synthase (iNOS) makes a great contribution to host defense and inflamma-tion. In many settings, lipopolysaccharide (LPS) induces iNOS expression through activation of the inhibitor of KB- α (IKB-α) -nuclear factor-KB (NF-KB) cascade, whereas interferon-γ (IFN-γ) acts through Janus kinase ( JAK)- signal transducer and activator of transcription 1 ( STAT1 ) signals. Heat shock factor 1 ( HSF1 ), a major regulator of heat shock protein transcription, has been shown to regulate the production of pro-inflammatory cytokines such as tumor necrosis factor-α(TNF-α) and interleukin-6 (IL-6). But it remains obscure whether and how HSF1 affects iNOS induction. Methods Western blot was used to measure the protein expression. The mRNA level was meas- ured by real time-PCR. Silence of HSF1 was achieved by small interfering RNA. Nitric oxide (NO) content and NF-KB binding activity were assayed by commercial kits. Chromatin immunoprecipitation (CHIP) was used to measure the binding activity of NF-KB and STAT1 to iNOS promoters. Results HSF1 inhibition or knockdown pre- vented the LPS- and/or IFN-γ-stimulated iNOS protein expression in cultured microglia. HSF1 inhibition blocked iNOS mRNA transcription. These inhibitory effects of HSF1 inhibition on iNOS expression were confirmed in brain tissues from endotoxemic mice. Further analysis showed that HSF1 inhibition had no effect on IKB-α degradation and NF-KB or STAT1 phosphorylation in LPS/IFN-γ-stimulated cells. The nuclear transport of active NF-KB or STAT1 was also not affected by HSF1 inhibition. But HSF1 inhibition reduced the binding of NF-KB and STAT1 to their DNA elements. In addition, HSF1 inhibition reduced NF-KB and STAT1 bindings to iNOS promoter inside the LPS/IFN-γ-stimulated cells. Conclusions This preventing effect of HSF1 inhibition on iNOS mRNA transcription presents the necessary role of HSF1 in iNOS induction. 展开更多
关键词 heat shock factor 1 lipopolysaccharide interferon--y INDUCIBLE NITRIC oxide SYNTHASE nuclear factor-KB signal transducer and ACTIVATOR of transcription 1
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Constitutively Expressed αB—Crystallin in Heat Schock Transcription Factor 1 Knockout Mice Myocardium
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作者 刘莉 张洪慧 +3 位作者 丁国宪 程蕴琳 晏良军 BENJAMINIvorJ 《Journal of Nanjing Medical University》 2003年第2期67-73,共7页
Objective-To investigate the effects of heat shock transcription factor 1) gene on the constitutivety expressed αB-CrystaUin (aBC) in mice myocardium. Methods-The expression levels of constitutive aBC in HSF1 knockou... Objective-To investigate the effects of heat shock transcription factor 1) gene on the constitutivety expressed αB-CrystaUin (aBC) in mice myocardium. Methods-The expression levels of constitutive aBC in HSF1 knockout (hsf1 - /- ) and HSFl wild type (As/1 + /+) mice myocardium were evaluated by western blot and immunohistochemistry. Results : The αBC levels in hsfl -/- and hsfl +/+ were 68. 42±4. 16, 100. 00±7. 58, respectively (P<0. 05, cytoso-lic fraction) , and 20. 53±1. 01, 37. 55±1. 91, respectively (P<0. 05, pellet fraction). The aBC signals decreased significantly in hsfl -/- myocardium when compared with those in hsfl +/+ myocardium stained with fluorescence immunohistochemistry. Conclusion-HSF1 is an important, but not the only factor, which mediates the constitutively expressed aBC. 展开更多
关键词 heat shock transcription factor 1 gene CONSTITUTIVE αB-Crystallin KNOCKOUT gene expression
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热激转录因子HSF调控植物非生物胁迫响应的作用机制
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作者 伍国强 张佳乐 魏明 《中国草地学报》 CSCD 北大核心 2024年第8期123-136,共14页
热激转录因子(Heat shock transcription factors,HSF)是植物抵抗热胁迫最重要的转录因子家族之一,广泛存在于真核生物。HSF拥有高度保守的DNA结合域,可参与复杂的胁迫信号转导和响应网络。HSF作为信号转导链的末端组分,介导非生物逆境... 热激转录因子(Heat shock transcription factors,HSF)是植物抵抗热胁迫最重要的转录因子家族之一,广泛存在于真核生物。HSF拥有高度保守的DNA结合域,可参与复杂的胁迫信号转导和响应网络。HSF作为信号转导链的末端组分,介导非生物逆境胁迫下靶标基因的表达。在逆境胁迫响应中,HSF受上游蛋白激酶介导磷酸化和泛素化或通过ABA信号通路结合热激元件,调控热激蛋白等下游基因的表达,从而提高植物的抗逆性。本文综述HSF的发现、结构、分类、调控机制及其在植物响应非生物逆境胁迫(极端温度、干旱、高盐碱和重金属等)中的作用机制,并对未来研究方向加以展望,以期为农作物和牧草抗逆性遗传改良提供理论依据和基因资源。 展开更多
关键词 热激转录因子 非生物逆境胁迫 调控机制 磷酸化 热激元件
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Comparative transcriptome analysis of the effect of different heat shock periods on the unfertilized ovule in watermelon(Citrullus lanatus) 被引量:2
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作者 ZHU Ying-chun SUN De-xi +5 位作者 DENG Yun AN Guo-lin LI Wei-hua SI Wen-jing LIU Jun-pu SUN Xiao-wu 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2020年第2期528-540,共13页
In vitro gynogenesis is an important tool used in haploid or homozygous double-haploid plant breeding.However,because of low repeatability,embryoid induction rate and quality,the molecular mechanisms remain poorly und... In vitro gynogenesis is an important tool used in haploid or homozygous double-haploid plant breeding.However,because of low repeatability,embryoid induction rate and quality,the molecular mechanisms remain poorly understood.Heat shock treatment can promote the transformation of the gametophytic pathway into the sporophyte pathway,which induces the occurrence of haploid.In this study,unfertilized ovaries were heat shocked for 0 h(A0)before flowering and for 0 h(A1),4 h(A3),8 h(A5),12 h(A7),and 24 h(A8),respectively,at 37℃at the first day of the flowering stage.The ovule enlargement rate was increased from 0%at 25℃to 96.8%at 37℃(24 h treatment).Thus,we aimed to investigate the gene expression patterns in unfertilized ovules of watermelon after different periods of heat shock by using RNA-Seq technology.The results showed that compared with A3,A5,A7,and A8,the biosynthesis of amino acid,glycine,serine and threonine metabolic pathways in A1 has changed significantly.This indicated that heat shock treatment affected the synthesis and transformation of amino acids during ovule expansion.The transcriptome data suggested gene expressions of ovule growth were significantly changed by heat-specific influences.The results provide new information on the complex relationship between in vitro gynogenesis and temperature.This provides a basis for further study of the mechanism of heat shock affecting the expansion of watermelon ovule. 展开更多
关键词 WATERMELON heat shock unfertilized ovule ovule enlargement transcription factors
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水稻热激转录因子HsfA2b调控非生物胁迫抗性的功能分析 被引量:1
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作者 邹修为 岳佳妮 +2 位作者 李志宇 戴良英 李魏 《生物技术通报》 CAS CSCD 北大核心 2024年第2期90-98,共9页
【目的】水稻在生长发育过程中,常会受到各种非生物胁迫而严重影响产量。热激转录因子作为植物抗逆过程中的一个重要元件,通过调控一系列胁迫响应基因的表达以提高植物抗逆性。探究水稻热激转录因子OsHsfA2b调控非生物胁迫的功能与初步... 【目的】水稻在生长发育过程中,常会受到各种非生物胁迫而严重影响产量。热激转录因子作为植物抗逆过程中的一个重要元件,通过调控一系列胁迫响应基因的表达以提高植物抗逆性。探究水稻热激转录因子OsHsfA2b调控非生物胁迫的功能与初步机理,为培育水稻抗逆新品种提供了优异基因资源和理论支撑。【方法】通过构建OsHsfA2b过量表达和RNAi的转基因水稻,分别观察水稻幼苗在高温、低温、干旱和高盐处理后的抗逆表型,统计存活率。同时,在逆境胁迫处理后,通过DAB染色检测水稻叶片活性氧(ROS)含量及RT-qPCR检测抗氧化途径相关基因OsSOD和OsCAT的表达量,分析OsHsfA2b对抗氧化途径的调控作用。【结果】在高温、低温、干旱和高盐等非生物胁迫条件下,水稻热激转录因子OsHsfA2b被显著诱导表达。与野生型NPB相比,OsHsfA2b过量表达转基因水稻对非生物胁迫的抗性明显增强,植株损伤程度较轻,存活率提高。相反,OsHsfA2b-RNAi水稻对非生物胁迫更加敏感,植株损伤严重,存活率降低。此外,在非生物胁迫条件下,OsHsfA2b过量表达转基因水稻比NPB和OsHsfA2bRNAi水稻体内活性氧的含量减少,并且OsHsfA2b显著诱导了抗氧化途径相关基因OsSOD和OsCAT的表达,OsHsfA2b参与抑制水稻体内活性氧的积累以降低逆境胁迫诱导的活性氧大量积累对植株的伤害。【结论】非生物胁迫下,水稻热激转录因子OsHsfA2b被显著诱导表达,可通过抗氧化途径正调控水稻对逆境胁迫的抗性。 展开更多
关键词 水稻 hsfA2b 热激转录因子 非生物胁迫 抗性 活性氧
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木薯MeHsfB3b转录因子与MeFKBP20蛋白互作关系验证
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作者 王超群 李琳琳 +3 位作者 陈银华 张肖飞 姚远 耿梦婷 《热带作物学报》 CSCD 北大核心 2024年第3期450-458,共9页
热激转录因子MeHsfB3b是调控木薯抗细菌性枯萎病的重要节点,前期通过酵母双杂交技术获得MeHsfB3b的候选互作蛋白MeFKBP20,该蛋白属于FKBP型肽脯氨酰顺反异构酶基因家族。本研究克隆获得MeFKBP20基因全长为561 bp,编码186个氨基酸,蛋白... 热激转录因子MeHsfB3b是调控木薯抗细菌性枯萎病的重要节点,前期通过酵母双杂交技术获得MeHsfB3b的候选互作蛋白MeFKBP20,该蛋白属于FKBP型肽脯氨酰顺反异构酶基因家族。本研究克隆获得MeFKBP20基因全长为561 bp,编码186个氨基酸,蛋白质分子质量为19.99 kDa,具有FKPB_C结构域。表达模式分析发现:MeFKBP20基因在木薯成熟叶片及根部高表达,在幼叶和叶柄的表达量较低;并能够迅速响应病原菌XpmCHN11诱导,持续保持较高的表达丰度,推测其参与了木薯响应病原菌侵染的过程。利用酵母双杂交点对点、双分子荧光互补实验证明MeHsfB3b蛋白通过在201~241 aa区域与MeFKBP20蛋白作用。本研究结果有助于进一步解析MeHsfB3b基因调控木薯对细菌性枯萎病的抗病机理。 展开更多
关键词 热激转录因子 FKBP型肽脯氨酰顺反异构酶 酵母双杂交 双分子荧光互补
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A Review of Heat Shock Proteins Research on Bemisia tabaci
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作者 Shunxiao Liu Kui Wang Vlasenko Volodymyr 《Agricultural Sciences》 2022年第3期393-403,共11页
Bemisia tabaci (Gennadius) (Homoptera: Aleyrodidae) is the most destructive invasive pests in agricultural production and has a high tolerance to heat. Heat shock proteins play an essential role in life activities suc... Bemisia tabaci (Gennadius) (Homoptera: Aleyrodidae) is the most destructive invasive pests in agricultural production and has a high tolerance to heat. Heat shock proteins play an essential role in life activities such as growth and development, reproduction and diapause of B. tabaci. At the same time, they are also crucial in resisting adverse environments and in adaptive evolution. The expression of heat shock protein in B. tabaci is not only related to temperature, but also to the tolerance of the environment. After receiving external stimuli, the expression level can be increased or decreased to maintain the stability of cells in vivo. This paper reviews the classification, biological characteristics, biological functions, and research status of HSPs in recent years. This mini-review will provide helpful information related to the use of heat shock proteins to study the occurrence and damage of B. tabaci. This has important theoretical and practical significance for revealing Hsps in explaining the population expansion mechanism of B. tabaci invasion and predicting population dynamics. 展开更多
关键词 Bemisia tabaci heat shock Proteins Molecular Chaperone heat shock transcriptional factor STRESS
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Genetic variation in a heat shock transcription factor modulates cold tolerance in maize
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作者 Lei Gao Lingling Pan +15 位作者 Yiting Shi Rong Zeng Minze Li Zhuoyang Li Xuan Zhang Xiaoming Zhao Xinru Gong Wei Huang Xiaohong Yang Jinsheng Lai Jianru Zuo Zhizhong Gong Xiqing Wang Weiwei Jin Zhaobin Dong Shuhua Yang 《Molecular Plant》 SCIE CSCD 2024年第9期1423-1438,共16页
Understanding how maize(Zea mays)responds to cold stress is crucial for facilitating breeding programs of cold-tolerant varieties.Despite extensive utilization of the genome-wide association study(GWAs)approach for ex... Understanding how maize(Zea mays)responds to cold stress is crucial for facilitating breeding programs of cold-tolerant varieties.Despite extensive utilization of the genome-wide association study(GWAs)approach for exploring favorable natural alleles associated with maize cold tolerance,few studies have successfully identified candidate genes that contribute to maize cold tolerance.In this study,we used a diverse panel of inbred maize lines collected from different germplasm sources to perform a GWAS on var-iations in the relative injured area of maize true leaves during cold stress-a trait very closely correlated with maize cold tolerance.We identified HsF21,which encodes a B-class heat shock transcription factor(HSF)that positively regulates cold tolerance at both the seedling and germination stages.Natural varia-tions in the promoter of the cold-tolerant HSF21Hap1 allele led to increased HSF21 expression under cold stress by inhibiting binding of the basic leucine zipper bziP68 transcription factor,a negative regulator of cold tolerance.By integrating transcriptome deep sequencing,DNA affinity purification sequencing,and targeted lipidomic analysis,we revealed the function of HsF21 in regulating lipid metabolism homeo-stasis to modulate cold tolerance in maize.In addition,we found that HsF21 confers maize cold tolerance without incurring yield penalties.Collectively,this study establishes HsF21 as a key regulator that en-hances cold tolerance in maize,providing valuable genetic resources for breeding of cold-tolerant maize varieties. 展开更多
关键词 heat shock factor hsf21 cold tolerance natural variation bZiP68 lipid metabolism MAIZE
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GmHSFA1基因克隆及其过量表达提高转基因大豆的耐热性 被引量:22
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作者 陈晓军 叶春江 +6 位作者 吕慧颖 徐民新 李葳 张利明 王超 罗淑萍 朱保葛 《遗传》 CAS CSCD 北大核心 2006年第11期1411-1420,共10页
热激转录因子在调节植物对逆境胁迫应答和热激蛋白基因表达方面起重要作用。采用生物信息学和比较基因组学方法结合RACE技术从大豆基因组中克隆到一个新的热激转录因子基因GmHsfA1,其cDNA全长1 781 bp,包含1个1 533 bp的开放阅读框,编... 热激转录因子在调节植物对逆境胁迫应答和热激蛋白基因表达方面起重要作用。采用生物信息学和比较基因组学方法结合RACE技术从大豆基因组中克隆到一个新的热激转录因子基因GmHsfA1,其cDNA全长1 781 bp,包含1个1 533 bp的开放阅读框,编码含有510个氨基酸残基的蛋白质(GenBank登录号为AY458843)。与其他转录因子的分子结构相似,GmHSFA1也含有4个典型的结构功能域-DNA结合域、寡聚域、核定位信号和C端激活域。BLAST分析表明,GmHSFA1与其同源性最高的番茄热激转录因子LpHSFA1之间的氨基酸序列相似性为52.46%。RT-PCR、Northern和遗传转化结果显示:1)GmHsfA1在大豆的不同组织中呈现组成型表达模式;2)常温下转基因大豆植株的GmHsfA1表达水平明显高于非转基因对照;3)GmHsfA1的过量表达激活了转基因大豆植株中热激蛋白基因GmHsp22在非诱导条件下的转录,并加强了高温胁迫下另2个热激蛋白基因GmHsp23和GmHsp70的表达;4)转GmHsfA1大豆植株的耐热温度(达52℃)明显高于非转基因植株。上述结果说明,GmHsfA1的过量表达激活或促进其下游3个热激蛋白基因的转录或表达,明显提高了转基因大豆植株的耐热能力。 展开更多
关键词 大豆 热激转录因子A1(hsfA1) 转基因 热胁迫 过量表达 耐热性
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HSF1和4-HNE的过度表达对前列腺癌恶性表型的影响 被引量:12
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作者 宋旭 王蓉 +3 位作者 张圣熙 龚敏 张耘 黄锦阳 《中华男科学杂志》 CAS CSCD 北大核心 2020年第8期722-725,共4页
目的:探讨热休克转录因子1(HSF1)和四羟基壬烯(4-HNE)在前列腺癌组织中的表达及其临床意义。方法:采用免疫组化方法检测50例前列腺癌组织及50例良性前列腺增生组织中HSF1和4-HNE的表达,分析HSF1和4-HNE的表达与Gleason分级之间的关系。... 目的:探讨热休克转录因子1(HSF1)和四羟基壬烯(4-HNE)在前列腺癌组织中的表达及其临床意义。方法:采用免疫组化方法检测50例前列腺癌组织及50例良性前列腺增生组织中HSF1和4-HNE的表达,分析HSF1和4-HNE的表达与Gleason分级之间的关系。结果:HSF1在良性前列腺增生组织中阳性表达率较低,为4.0%。50例前列腺癌组织中HSF1阳性表达率在高、中、低分化癌中分别为37.5%、50.0%、75.0%,呈明显上升趋势,两组HSF1阳性表达差异有统计学意义(P=0.001)。HSF1的阳性表达率在不同Gleason分级间差异有统计学意义(P=0.025)。4-HNE在良性前列腺增生组织中阳性表达率较低,仅为6.0%。50例前列腺癌组织中4-HNE阳性表达率在高、中、低分化癌中分别为81.3%、92.9%、100.0%,呈上升趋势,两组4-HNE表达的差异有统计学意义(P=0.001)。4-HNE的阳性表达率在不同Gleason分级间差异有统计学意义(P=0.029)。前列腺癌组织中HSF1和4-HNE阳性表达呈正相关(r=0.947,P=0.001)结论:HSF1和4-HNE的过度表达与前列腺癌的分级相关,可以作为判断前列腺癌恶性程度具有重要参考价值的新生物学标记物。 展开更多
关键词 前列腺肿瘤 热休克转录因子1 四羟基壬烯 免疫组化
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苹果Hsf家族成员的序列特征、表达与进化分析 被引量:3
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作者 张国俊 王婷婷 +2 位作者 胡利宗 李书粉 高武军 《华北农学报》 CSCD 北大核心 2017年第2期71-80,共10页
为全面了解苹果基因组中热激转录因子(Hsf)的序列特征及进化,采用生物信息学手段,在苹果全基因组水平上鉴定出50个MdHsf基因,并对其系统发育关系、序列特征、表达情况以及选择压力进行详细分析。系统发育与序列分析显示:与拟南芥和水稻... 为全面了解苹果基因组中热激转录因子(Hsf)的序列特征及进化,采用生物信息学手段,在苹果全基因组水平上鉴定出50个MdHsf基因,并对其系统发育关系、序列特征、表达情况以及选择压力进行详细分析。系统发育与序列分析显示:与拟南芥和水稻相似,50个MdHsf基因可分为A、B、C 3个亚族;2个或多个MdHsf基因位于同一个末端进化支,说明该基因家族在苹果中发生了物种特异性扩增;尽管MdHsf基因的内含子数目和长度变异较大,但其蛋白的保守基序和功能结构域具有较高的保守性,这可能与功能约束有关。基于EST数目,可推知:除了MdHsfA2a和MdHsfA3a/b/c等14个基因没有相应的EST外,其余72%的基因都有转录活性。选择压力检测和结构建模分析显示:在36个MdHsf蛋白的选择压力检测中,位点模型未鉴定到正选择位点的存在;而在显著水平下(P<0.05),分支-位点模型在d和e进化分支上,共检测到5个正选择位点,它们是28R、30L、35D、51M、67V,其中28R和30L位于Hsf结构域中,35D、51M和67V位于Hsf结构域之外,这说明除了MdHsfA4d/e和MdHsf C1a/b发生快速进化外,其他成员受控于纯净选择,具有高度保守性。综合以上研究结果,苹果基因组中存在多种热激转录因子,其蛋白的保守基序和功能结构域具有较高保守性,大多具有转录活性,在进化上该家族受纯净选择主导。 展开更多
关键词 苹果 热激转录因子 表达 进化
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松材线虫热激转录因子Bx-HSF-1基因的克隆及表达分析 被引量:5
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作者 王峰 马玲 +4 位作者 陈俏丽 王博文 郝欣 何芳林 柳燕 《林业科学》 EI CAS CSCD 北大核心 2016年第12期92-98,共7页
【目的】通过对热激转录因子基因Bx-HSF-1的克隆和表达分析,为深入研究c GMP、TGFβ-like和Insulin/IGF等热激抗逆代谢路径提供理论基础。【方法】根据松材线虫基因组数据设计引物克隆松材线虫Bx-HSF-1基因。进一步对Bx-HSF-1进行Gene O... 【目的】通过对热激转录因子基因Bx-HSF-1的克隆和表达分析,为深入研究c GMP、TGFβ-like和Insulin/IGF等热激抗逆代谢路径提供理论基础。【方法】根据松材线虫基因组数据设计引物克隆松材线虫Bx-HSF-1基因。进一步对Bx-HSF-1进行Gene Ontology基因功能预测、系统发育分析、保守结构域分析等生物信息学分析。应用JASPAR预测Bx-HSF-1靶位点DNA序列,根据松材线虫基因组数据筛选含有该序列的靶基因,并通过蛋白质三维结构分析和分子对接验证。应用荧光定量q-PCR检测Bx-HSF-1及其靶基因在热激和低温处理条件下4个时间点的表达量,结合转录组数据对Bx-HSF-1及其靶基因的表达量相关性进行分析。采用原位杂交对Bx-HSF-1基因在松材线虫体内的表达部位进行了分析。【结果】根据基因组数据,克隆到松材线虫热激转录因子Bx-HSF-1基因长度为1 404 bp的涵盖完整蛋白编码区基因序列。结构域分析表明Bx-HSF-1基因编码蛋白质的氨基酸序列含有DNA结合结构域和亮氨酸拉链结构域。GO分析表明该基因参与延长线虫寿命、正向调控繁殖率、运动、热激反应、抗逆态幼虫发育和抗逆反应等生理过程,并具有特异DNA序列结合的分子功能。氨基酸疏水性分析表明该DNA结合结构域由3个α螺旋和4个反向平行的β折叠构成一个疏水核心,该疏水核心可以与热激元件靶位点互作。对松材线虫基因组进行BLAST比对筛选出Bx-DAF-7是Bx-HSF-1的靶基因,Bx-DAF-7基因TATA盒上游序列具有Bx-HSF-1靶位点序列。荧光定量q-PCR结果表明热激和低温处理同样促进Bx-HSF-1转录、抑制Bx-DAF-7转录,Bx-HSF-1与Bx-DAF-7的表达量负相关。原位杂交结果表明Bx-HSF-1基因在正常培养线虫体内各细胞均有微量表达,热激培养线虫前体部信号显著,峡部和肠区体壁信号较显著。【结论】本研究表明Bx-HSF-1具有特异DNA序列结合的分子功能。预测抗逆态幼虫形成基因Bx-DAF-7是Bx-HSF-1靶基因。转录组测序及q-PCR结果表明Bx-HSF-1与Bx-DAF-7的表达量负相关。Bx-HSF-1基因受温度刺激后多数体细胞内均有较高表达,特别是神经元、体壁肌肉细胞和皮下细胞中表达显著。 展开更多
关键词 松材线虫 热激转录因子 热激抗逆 低温处理
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转GmHSFA1基因大豆的耐热性评价 被引量:4
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作者 吴广锡 刘丽君 +4 位作者 杨德光 唐晓飞 魏崃 王鹏飞 王兴宇 《大豆科学》 CAS CSCD 北大核心 2012年第3期341-346,共6页
在高温胁迫条件下对11个转GmHSFA1基因的大豆品系及其受体的GmHSFA1及靶基因GmHSP70、GmHSP22和GmHSP17.9的表达和生理生化、光合特性及产量性状变化进行了研究,并采用胁迫系数与灰色关联分析相结合方法,对转基因大豆材料耐高温能力进... 在高温胁迫条件下对11个转GmHSFA1基因的大豆品系及其受体的GmHSFA1及靶基因GmHSP70、GmHSP22和GmHSP17.9的表达和生理生化、光合特性及产量性状变化进行了研究,并采用胁迫系数与灰色关联分析相结合方法,对转基因大豆材料耐高温能力进行综合评价。结果表明:多数转基因大豆品系在常温和热诱导条件下GmHSFA1及靶基因GmHSP70、GmHSP22和GmHSP17.9表达水平明显高于非转基因受体;高温处理后,绝大多数转基因材料的脯氨酸含量增幅都要明显高于非转基因受体,丙二醛含量除HTH-4以外增幅均小于非转基因受体,可溶性糖含量增幅全部高于非转基因受体材料。多数转基因材料的光合性状与产量性状降幅均小于非转基因受体。关联度分析表明与非转基因受体相比,有9个转GmHSFA1基因品系的耐热能力得到明显提高。 展开更多
关键词 大豆 转基因 热激转录因子 热激蛋白 热胁迫
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HSF1基因敲除小鼠胚胎成纤维细胞的永生化 被引量:6
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作者 刘梅冬 张华莉 +4 位作者 龚环宇 陈广文 王慷慨 鄂顺梅 肖献忠 《中南大学学报(医学版)》 CAS CSCD 北大核心 2006年第2期174-177,共4页
目的:建立HSF1-/-,HSF1+/+两种基因型小鼠胚胎成纤维永生化细胞系,为HSF1的功能研究提供实验模型。方法:用脂质体介导的基因转染法将pSV 3 neo质粒导入HSF1-/-,HSF1+/+两种基因型小鼠胚胎成纤维细胞,经G 4 1 8筛选,抗性克隆扩大培养,建... 目的:建立HSF1-/-,HSF1+/+两种基因型小鼠胚胎成纤维永生化细胞系,为HSF1的功能研究提供实验模型。方法:用脂质体介导的基因转染法将pSV 3 neo质粒导入HSF1-/-,HSF1+/+两种基因型小鼠胚胎成纤维细胞,经G 4 1 8筛选,抗性克隆扩大培养,建立永生化细胞系;用PCR检测两种细胞株中目的基因的整合,用RT-PCR法鉴定SV 4 0T基因在转染细胞中的表达;用W estern b lot检测所建细胞株的诱导型热休克蛋白7 0的表达情况。结果:有3个细胞克隆已扩大培养稳定传代达6个月,经鉴定SV 4 0 T抗原已整合到两种细胞中且稳定表达,HSF1-/-胚胎成纤维细胞热休克蛋白7 0的诱导表达消失。结论:成功建立永生化HSF1-/-,HSF1+/+两种基因型小鼠胚胎成纤维细胞。 展开更多
关键词 hsf1 基因敲除 胚胎成纤维细胞 永生化 SV40 T抗原
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大豆热激转录因子GmHsFA1对高温和干旱信号的表达反应 被引量:5
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作者 吴广锡 魏崃 +4 位作者 唐晓飞 王伟威 王鹏飞 王兴宇 刘丽君 《分子植物育种》 CAS CSCD 北大核心 2013年第1期62-71,共10页
本研究将热激转录因子GmHsFA1基因构建在植物表达载体pCAMBIA3300中,以大豆子叶节作为受体,通过农杆菌介导法将热激转录因子GmHsFA1基因导入到高产品种黑农53中,获得一批转基因株系。采用Real-time PCR的方法对各代株系GmHsFA1基因的转... 本研究将热激转录因子GmHsFA1基因构建在植物表达载体pCAMBIA3300中,以大豆子叶节作为受体,通过农杆菌介导法将热激转录因子GmHsFA1基因导入到高产品种黑农53中,获得一批转基因株系。采用Real-time PCR的方法对各代株系GmHsFA1基因的转录水平进行相对定量分析,确定转基因大豆中热激转录因子GmHsFA1的表达量明显提高。利用转热激转录因子GmHsFA1大豆研究了高温和干旱胁迫下,大豆热激转录因子GmHsFA1应对高温和干旱信号的基因表达,生理生化特性,光合特性及产量性状的变化反应,采用胁迫系数与灰色关联分析相结合方法,对转GmHsFA1基因大豆的耐热性和抗旱性进行综合鉴定和评价。结果表明:转热激转录因子GmHsFA1大豆品系在高温和干旱诱导条件下,大豆叶片中GmHsFA1、GmHSP70、GmHSP22和GmHSP17.9基因的表达量明显高于非转基因受体黑农53,叶肉细胞中的脯氨酸含量和可溶性糖含量明显高于受体,丙二醛含量增幅较小,叶片的光合特性受胁迫变化较小,光合特性能力和产量性状表现高于受体,表明过量表达的热激转录因子GmHsFA1大豆植株的耐热能力和抗干旱能力得到明显的提高。 展开更多
关键词 大豆 热激转录因子GmhsfA1 农杆菌介导 转基因 耐热性 抗旱性
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HSF1与XAF1基因在子宫内膜癌组织中的表达及其相关性分析 被引量:9
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作者 周卓 李佳 +3 位作者 张千锋 张建芳 王建 陈必良 《西安交通大学学报(医学版)》 CAS CSCD 北大核心 2015年第2期231-234,共4页
目的检测HSF1与XAF1基因在子宫内膜组织中的表达,分析其与临床病理学特征的关系。方法采用免疫组织化学S-P法检测64例子宫内膜癌组织(EC组)、33例正常子宫内膜组织(NE组)中两者的表达情况,并观察其相关性。结果子宫内膜癌组织中HSF1阳... 目的检测HSF1与XAF1基因在子宫内膜组织中的表达,分析其与临床病理学特征的关系。方法采用免疫组织化学S-P法检测64例子宫内膜癌组织(EC组)、33例正常子宫内膜组织(NE组)中两者的表达情况,并观察其相关性。结果子宫内膜癌组织中HSF1阳性表达率为76.6%,显著高于正常子宫内膜组织36.4%(P<0.05);子宫内膜癌及正常子宫内膜组织XAF1阳性率分别为31.2%和72.7%,差别具有统计学意义(P<0.05);在EC组中HSF1的阳性表达率在不同的组织学分级、肌层浸润及淋巴结转移间差异有统计学意义(P<0.05);在EC组中XAF1的阳性表达率在不同的组织学分级、肌层浸润、手术病理分期及淋巴结转移间差异有统计学意义(P<0.05);子宫内膜癌组织中HSF1与XAF1表达水平呈负相关(P<0.05)。结论在子宫内膜癌组织中HSF1的高表达可能通过抑制XAF1的表达致使细胞过度生长,抑制肿瘤细胞发生凋亡,导致肿瘤进一步发展。 展开更多
关键词 子宫内膜癌 热休克转录因子 X-染色体相关凋亡抑制因子 细胞凋亡
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