共查询到20条相似文献，搜索用时 50 毫秒
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拟南芥RPS14(Ribosomal protein S14)是从拟南芥中分离出的一种核糖体蛋白,是核糖体40S亚基的组成成分,属于S11P核糖体蛋白家族,主要负责RNA转录后加工。提取拟南芥总RNA,用RT-PCR技术得到RPS14全长基因,经T-A克隆插入到pEASY-T3载体上。利用亚克隆法成功构建pGEX-6P-1-RPS14原核表达质粒。GST-RPS14融合蛋白在大肠杆菌BL21(DE3)中表达,分子量约为43 kD。运用同样方法构建pET-28a-AK6原核表达质粒,获得大小约为26 kD的HIS-AK6融合蛋白。运用体外pull-down技术,并用SDS-PAGE和Western blot进行验证,证明拟南芥RPS14蛋白与AK6蛋白之间存在相互作用。因此,推测拟南芥的AK6与RPS14蛋白共同参与核酸代谢过程,影响RNA转录后加工,进而抑制拟南芥茎细胞的生长,使植株表现矮小特征。 相似文献
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以模式植物拟南芥为材料,采用RT-PCR分析、基因克隆、转化等方法对CIPK1基因功能进行了研究.结果发现,CIPK1(CBL-interacting protein kinase 1)基因在拟南芥茎和花中表达量最高,在叶中表达量最低;ABA能迅速诱导CIPK1基因的表达,GA则抑制该基因的表达,但2,4-D和6-BA对该基因的表达无明显影响;通过对CIPK1基因转基因突变体进行ABA处理,发现转基因拟南芥种子的萌发率明显高于野生型.以上结果表明CIPK1基因的表达具有组织特异性,并且该基因参与ABA和GA信号传导,尤其在ABA促进种子休眠的信号传导途径中具有非常重要的作用. 相似文献
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拟南芥、荠菜DREB1A基因的克隆与植物表达载体的构建 总被引：2，自引：0，他引：2
用PCR方法从拟南芥和荠菜中分别克隆了DREB1A基因.序列分析发现从拟南芥中克隆的AtDREB1A基因与已发表的AtDREB1A基因序列(DQ372533)的同源性为99.69%.首次从荠茱中克隆的CbDREB1A基因序列(EF156749)与DQ372533的同源性为99.54%.利用这两个基因分别构建了两个诱导表达载体(prd29A/AtDREB1A,prd29A/CbDREB1A)和两个组成型表达载体(pCaMV35S/AtDILEBlA,pCaMV35S/CbDREB1A),以便进一步开展植物抗旱基因工程研究. 相似文献
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Yptl蛋白是酵母唯一的Rab1 GTP酶,调控囊泡从内质网到高尔基体的运输.酵母温敏突变株 ASY01是一个Ypt1基因功能部分缺失菌株,在26℃可以正常生长,但在37℃不能生长.拟南芥有4个Rab1基因,分别是AtRab1A1、AtRab1B1、AtRab1B2、AtRab1C1.克隆了所有4个AtRab1基因,构建酵母表达载体,转化温敏突变型酵母ASY01.温度敏感性实验结果表明,所有转基因菌株在37℃都恢复正常生长.说明拟南芥4个Rab1基因都与酵母Ypt1基因功能互补,都具有调节囊泡从内质网到高尔基体运输的功能. 相似文献
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林浴霞;周燕;刘林川 《植物科学学报》2025,(6):749-757
生长素是调控植物生长发育的重要激素,TWD1(Twisted dwarf 1)介导生长素转运蛋白在分泌途径中的折叠。本研究利用生物信息学和分子生物学手段从黄梁木（Neolamarckia cadamba(Roxb.) Bosser）中鉴定并克隆了一个与拟南芥（Arabidopsis thaliana(L.) Heynh.）AtTWD1同源的基因,将其命名为NcTWD1。分析结果显示,NcTWD1编码序列长度为1 098 bp,编码365个氨基酸。NcTWD1和AtTWD1蛋白质三维空间结构相似,均含有FKBP结构域、TRP结构域和C端膜锚定结构域。RT-qPCR分析结果表明,NcTWD1在黄梁木的根、茎、芽尖、嫩叶、老叶、叶柄和幼苗中均有表达。亚细胞定位分析结果显示NcTWD1主要定位于内质网。本研究进一步构建过表达载体,发现在拟南芥twd1-4突变体中异源过量表达NcTWD1能够恢复其矮小卷曲的表型。利用免疫共沉淀技术分析,发现黄梁木NcTWD1与生长素转运蛋白NcABCB1存在相互作用,表明NcTWD1在黄梁木中具有和拟南芥AtTWD1相似的生物学功能。 相似文献
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前期研究表明ABL1可能在植物叶发育过程中扮演重要的角色,其突变表现为叶片生长迟缓、成熟叶片叶缘缺刻明显等生长缺陷特征。该研究利用图位克隆及其精细定位技术,将ABL1基因锁定在2个SSLP标记T23K8和T8F5之间,该区间包含44个基因。通过生物信息学成功找到ABL1突变基因为拟南芥FAS1,该基因编码染色质组装因子CAF1的一个亚基,在植物顶端分生组织生长调控中扮演重要角色。RT-PCR结果显示,该基因表达受阻,功能互补实验证实abl1突变体的确是FAS1基因的一个新等位突变。研究结果暗示,ABL1/FAS1在植物叶形态建成中也起着重要作用。 相似文献
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Gorou Horiguchi Nathalie Gonzalez Gerrit T.S. Beemster Dirk Inzé Hirokazu Tsukaya 《The Plant journal : for cell and molecular biology》2009,60(1):122-133
The number of cells in an organ is a major factor that specifies its size. However, the genetic basis of cell number determination is not well understood. To obtain insight into this genetic basis, three grandifolia-D ( gra-D ) mutants of Arabidopsis thaliana were characterized that developed huge leaves with two to three times more cells than the wild-type. Genetic and microarray analyses showed that a large segmental duplication had occurred in all the gra-D mutants, consisting of the lower part of chromosome 4. In the duplications, genes were found that encode AINTEGUMENTA (ANT), a factor that extends the duration of cell proliferation, and CYCD3;1, a G1 /S cyclin. The expression levels of both genes increased and the duration of cell proliferation in the leaf primordia was extended in the gra-D mutants. Data obtained by RNAi-mediated knockdown of ANT expression suggested that ANT contributed to the huge-leaf phenotype, but that it was not the sole factor. Introduction of an extra genomic copy of CYCD3;1 into the wild-type partially mimicked the gra-D phenotype. Furthermore, combined elevated expression of ANT and CYCD3;1 enhanced cell proliferation in a cumulative fashion. These results indicate that the duration of cell proliferation in leaves is determined in part by the interaction of ANT and CYCD3;1 , and also demonstrate the potential usefulness of duplication mutants in the elucidation of genetic relationships that are difficult to uncover by standard single-gene mutations or gain-of-function analysis. We also discuss the potential effect of chromosomal duplication on evolution of organ size. 相似文献
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Planchais S Samland AK Murray JA 《The Plant journal : for cell and molecular biology》2004,38(4):616-625
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We aimed to investigate abiotic stress (WAK, HvPIP1.1, HvPIP1.2, HvPIP1.3, HvPIP1.5, CYCD3, DREB2) and brassinosteroid-related gene (DWARF4) expressions in barley (Hordeum vulgare L. cv. “Hilal”) roots grown under different salt (150 and 250 mM), HBR (0.5 and 1 μM), and salt + HBR applications during 48 and 72 h at dark with their controls. Phylogenetic trees were also constructed to observe relationships among genes found in other plants. The expression of HvPIP1.2 and WAK reduced after salt treatment while HvPIP1.3, DREB2 and DWARF4 expressions increased. HvPIP1.1, HvPIP1.2, HvPIP1.3, HvPIP1.5 and DWARF4 expressions were upregulated under only HBR applications. Salt + HBR treatments increased HvPIP1.1, DREB2 and DWARF4 but decreased HvPIP1.2. Phylogenetic analyses indicated that Oryza sativa L. shared similar sequences with HvPIP1.5. CYCD3 could diverge relatively earlier from cyclin genes during evolution as it segregates in a distinct clade. Sorghum bicolor showed sequence homology with DREB2. Oryza australiensis L. and DWARF4 were found in the same clade. To our knowledge, this is the first detailed report related to salt stress and HBR applications in terms of the expression of different genes in barley, providing a valuable information for molecular breeding improvement of stress-related traits. 相似文献
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G1到S期的转换是植物细胞周期中一个关键的调控点,而D型细胞周期蛋白(CYCD)在这一转换过程中起着重要作用.CYCD通过感受外界信号的刺激,调控细胞周期进程,进而影响植物的生长发育.为研究木本植物中不同CYCD基因家族的功能,从黑杨中克隆出6个CYCD基因,并将其转化至酵母G1期细胞周期蛋白突变体进行功能鉴定.各家族... 相似文献
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Gaia Bertolotti Simon Josef Unterholzner Daria Scintu Elena Salvi Noemi Svolacchia Riccardo Di Mambro Veronica Ruta Francisco Linhares Scaglia Paola Vittorioso Sabrina Sabatini Paolo Costantino Raffaele Dello Ioio 《Current biology : CB》2021,31(2):420-426.e6
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目的:为脂肪细胞分化提供数据,加深对3T3-L1细胞分化机制的了解.方法:应用AFM对3T3-L1前脂细胞的形貌、超微结构、机械性能和细胞骨架进行了可视化研究.结果:3T3-L1细胞舒展,伪足丰富,膜表面有大小不一的斑块和突起.通过统计分析得出3T3-L1细胞的高低差,均方根粗糙度、平均粗糙度和平均高度分别为622.3nm、77.34nm、55.80nm、393.1nm;AFM针尖与细胞膜表面的相对粘弹力为95.10±19.41pN,平均硬度为2.36±0.39mN/m,杨氏模量为4.85±0.99kPa.AFM对3T3-L1细胞骨架成像,观察到骨架南排列整齐的大纤维束和细小的微纤维以及颗粒状蛋白组成,形成网络结构.结论:细胞形貌结合细胞的机械性能可知3T3-L1细胞生长状态良好,细胞的移动迁移能力强. 相似文献
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成纤维细胞生长因子-21(FGF-21)是FGF家族的成员之一．近年发现FGF-21是一种新的代谢调节因子．从小鼠肝脏克隆FGF-21 cDNA,经测序正确后亚克隆至具有羟胺切割位点的小泛素相关修饰物表达载体上,转化宿主菌Rosetta,得到的转化子经IPTG诱导后获得稳定、高效、可溶的表达产物．表达产物经羟胺切割、透析、复性、柱层析纯化后,在每升宿主菌中可获得4 mg纯度为95%的成熟鼠源FGF-21蛋白,利用葡萄糖氧化酶-过氧化物酶(POD-GOD)法在小鼠3T3-L1脂肪细胞中进行生物学活性检测．结果表明,鼠源FGF-21具有促进脂肪细胞吸收葡萄糖的作用,短期作用(1 h)与胰岛素相似,长期作用(8和12 h)明显优于胰岛素．这一结果为以鼠源FGF-21为模型进一步研究FGF-21的生物学活性及其在糖代谢方面的作用机理奠定了基础． 相似文献
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转录因子AP-2是一个功能重要的基因家族,AP-2α,在协同调控脂肪细胞分化成熟,以及脂肪因子分泌的过程中发挥重要作用.为了阐明其协同调控的机制,本文采用anti-AP-2α抗体免疫共沉淀技术,从小鼠前脂肪细胞313-L1中分离AP-2α相互作用蛋白,然后用基质辅助激光解吸/电离飞行时间质谱结合数据库查询鉴定AP-2a结合蛋白,找到了aeyl-Coenzyme A dehydrogenase,long-chain、Protein-L-isoaspartate(D-aspartate)O-methyltransferase和T-complex protein 1 subunit epsilon三个与AP-2α相互作用的蛋白.为进一步研究小鼠的前脂肪细胞中AP-2α蛋白聚集及在糖脂代谢调控中可能的作用机制提供了重要依据. 相似文献
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Lei Jiang Xiao Qing Chen Ming Jing Gao Wai Lee Jie Zhou Yun Fu Zhao Guo Dong Wang 《Journal of cellular and molecular medicine》2019,23(4):2769-2781
Periodontitis, an oral inflammatory disease caused by periodontal pathogen infection, is the most prevalent chronic inflammatory disease and a major burden on healthcare. The TAM receptor tyrosine kinases (Tyro3, Axl and Mertk) and their ligands (Gas6 and Pros1) play a pivotal role in the resolution of inflammation and have been associated with chronic inflammatory and autoimmune diseases. In this study, we evaluated the effects of exogenous Pros1 in in vitro and in vivo models of periodontitis. We detected higher Pros1 but lower Tyro3 levels in inflamed gingival specimens of periodontitis patients compared with healthy controls. Moreover, Pros1 was mostly localized in the gingival epithelium of all specimens. In cultured human gingival epithelial cells (hGECs), Porphyromonas gingivalis LPS (p.g‐LPS) stimulation down‐regulated Pros1 and Tyro3. Exogenous Pros1 inhibited p.g‐LPS–induced production of TNF‐α, IL‐6, IL‐1β, MMP9/2 and RANKL in a Tyro3‐dependent manner as revealed by PCR, Western blot analysis, ELISA and gelatin zymography. Pros1 also restored Tyro3 expression down‐regulated by p.g‐LPS in hGECs. In rats treated with ligature and p.g‐LPS, administration of Pros1 attenuated periodontitis‐associated gingival inflammation and alveolar bone loss. Our mechanistic studies implicated SOCS1/3 and STAT1/3 as mediators of the in vitro and in vivo anti‐inflammatory effects of Pros1. Collectively, the findings from this work supported Pros1 as a novel anti‐inflammatory therapy for periodontitis. 相似文献