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清热活血健脾中药对大鼠肝癌基因转录差异的调整 被引量:12
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作者 方肇勤 管冬元 梁尚华 《世界华人消化杂志》 CAS 2003年第3期276-280,共5页
目的:观察中药抑癌扶正行气活血方(全方)及其不同治法的拆方:清热方、活血方、健脾方对肝癌大鼠的作用和对肝组织基因转录的整体调节作用.方法:采用二乙基亚硝胺(DEN)诱发大鼠肝癌,设正常组、模型组、不同中药组及FT-207组对照,分别观... 目的:观察中药抑癌扶正行气活血方(全方)及其不同治法的拆方:清热方、活血方、健脾方对肝癌大鼠的作用和对肝组织基因转录的整体调节作用.方法:采用二乙基亚硝胺(DEN)诱发大鼠肝癌,设正常组、模型组、不同中药组及FT-207组对照,分别观察各组大鼠生存率、肝、体、肝/体及肝组织的病理、甲胎蛋白(AFP)免疫组化变化;以及采用DDPCR技术显示正常肝组织与模型组肝癌组织中呈差异转录的cDNA片段,Northernblot验证这些cDNA片段在各组中转录的差异,并将呈差异转录的cDNA片段进行克隆与测序.结果:全方其及拆方均能不同程度地改善肝癌大鼠的一般情况,提高大鼠的生存率.各拆方中以清热组与活血组为优;全方其及拆方均能不同程度地抑制肿瘤的生长及肝脏AFP的合成,其中以全方组与清热组为优;采用DDPCR结合Northernblot筛选出在各组中呈显著差异转录的9个阳性cDNA片段,经与Genbank比较4个为新的基因,且这9个基因在肝癌发生与转归中的作用至今未见报道;全方及其拆方对这些基因的转录水平有不同程度的调节作用,其中DD29下调57-78%,DD11、DD25也有下调至60%和78%,使基因转录水平接近正常肝组织.结论:全方及其不同治法的拆方中药对大鼠肝癌具有直接治疗作用,能不同程度地广泛地调控肝组织(含肝癌组织)有关基因的转录水平. 展开更多
关键词 清热活血健脾中药 大鼠 肝癌 基因转录差异 调整
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基于转录组测序技术分析猪德尔塔冠状病毒感染ST细胞机制的初步研究 被引量:1
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作者 谢永生 张昆丽 +1 位作者 张建峰 贺东生 《中国预防兽医学报》 CAS CSCD 北大核心 2024年第3期236-244,共9页
猪德尔塔冠状病毒(PDCoV)感染可导致仔猪出现水样腹泻和死亡,严重危害猪的健康。为研究PDCo V感染引起宿主细胞基因转录水平的变化及初步研究其感染机制,本研究提取PDCo V感染组和不感染该病毒的阴性对照组ST细胞样品总RNA,构建二者的c... 猪德尔塔冠状病毒(PDCoV)感染可导致仔猪出现水样腹泻和死亡,严重危害猪的健康。为研究PDCo V感染引起宿主细胞基因转录水平的变化及初步研究其感染机制,本研究提取PDCo V感染组和不感染该病毒的阴性对照组ST细胞样品总RNA,构建二者的cDNA文库后采用Illumina HiSeq 6000高通量测序平台进行转录组测序(RNA-Seq)。结果显示,所有样品的测序错误率均≤0.03%;Q20/%和Q30/%分别达98%和94%以上;GC含量在51.09%~55.15%;相关性分析结果显示,阴性对照组及感染组细胞样品各组内的R值均>0.9,上述结果表明测序数据可靠,可用于后续转录组学数据分析。采用ggplot2软件分析两组细胞组内测序结果的相关性,并以相关系数R>0.9判定各组内测序数据的准确性;采用DESeq2软件并以P<0.05和log2(Fold change)>1为条件筛选两组细胞中转录显著差异基因;采用ClusterProfiler R软件对转录显著差异基因进行GO功能和KEGG信号通路的富集分析;采用Pheatmap软件对各组ST细胞中与免疫反应相关的转录显著差异基因进行聚类分析;随机选择9个转录显著差异基因经RT-q PCR验证RNA-Seq的结果。筛选结果显示,与阴性对照组细胞相比,共筛选出5719个转录显著差异基因,其中3573个转录显著上调基因,2146个转录显著下调基因。GO功能和KEGG信号通路富集分析结果显示,转录显著上调的基因与ST细胞的免疫应答、防御反应等功能有关;转录显著下调的基因与细胞的氧化还原和新陈代谢等功能有关;转录显著上调的基因参与细胞因子受体互作、天然免疫反应相关、肿瘤坏死因子等的信号通路;转录显著下调的基因与细胞物质代谢信号通路有关,包括氨基酸代谢、脂代谢、乙醛酸和二羧酸代谢等信号通路。与免疫反应相关的转录显著差异基因的聚类分析结果显示,与阴性对照组细胞相比,PDCo V感染的ST细胞中白细胞介素基因(IL-1R1、1L-6、IL-10RB、IL-15和IL-12A等)、抗病毒基因(MX1、OAS)和干扰素基因(INFAR1、IFN-ALPHAOMEGA)的转录水平均显著上调。上述结果表明PDCo V感染后,ST细胞的天然免疫反应被激活,但细胞的代谢反应降低,这更有利于减弱病毒在ST细胞中的复制,起到一定的抗病毒效果。9个转录显著差异基因的RT-q PCR结果与RNA-seq结果基本一致,表明转录组测序数据可靠。本研究为进一步解析PDCo V的感染机制提供参考依据。 展开更多
关键词 猪德尔塔冠状病毒 转录组测序 转录显著差异基因
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伪狂犬病病毒感染小鼠的背根神经节后天然免疫相关转录组测序及分析 被引量:3
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作者 王冰 吴红霞 +4 位作者 李淼 高莹 袁梦淇 仇华吉 孙元 《中国预防兽医学报》 CAS CSCD 北大核心 2022年第6期591-597,共7页
伪狂犬病病毒(PRV)的天然宿主是猪,但是能够感染多种哺乳动物。PRV是一种嗜神经性α疱疹病毒,小鼠感染PRV后会产生严重的神经症状和神经炎症。为了探究小鼠感染PRV后其背根神经节(DRG)的天然免疫应答情况,本研究将PRV TJ株以10^(6)pfu... 伪狂犬病病毒(PRV)的天然宿主是猪,但是能够感染多种哺乳动物。PRV是一种嗜神经性α疱疹病毒,小鼠感染PRV后会产生严重的神经症状和神经炎症。为了探究小鼠感染PRV后其背根神经节(DRG)的天然免疫应答情况,本研究将PRV TJ株以10^(6)pfu剂量接种小鼠,在濒死期剖杀小鼠,取其DRG进行转录组测序,并使用DESeq2软件对测序结果分析,结果显示,与对照组相比,PRV感染小鼠的DRG内共有6 502条基因的转录水平发生改变,其中转录水平升高的基因有3 703条,降低的基因有2 799条(差异倍数>2,且P≤0.05)。利用clusterProfiler R软件对发生差异转录的基因进行了KEGG分析,结果显示,转录水平发生差异的基因参与细胞内20种信号通路途径,其中12条信号通路与天然免疫反应有关,提示PRV感染小鼠后会激活DRG内广泛的天然免疫反应。差异转录基因数量最多的5条天然免疫信号通路分别是丝裂原活化蛋白激酶(MAPK)信号通路、核苷酸结合寡聚化结构域样受体(NLR)信号通路、肿瘤坏死因子(TNF)信号通路、细胞凋亡(Apoptosis)信号通路和信号转导及转录激活因子(JAK-STAT)信号通路,上述天然免疫反应通路可能在PRV诱导的神经炎症方面有重要作用。本研究随机选择了10条转录水平存在差异的基因,采用荧光定量PCR验证,结果显示其变化趋势均与转录组测序数据一致。本研究首次分析了PRV感染小鼠DRG内转录组的变化,为研究PRV感染后诱导宿主的天然免疫反应相关信号通路提供了参考,并为PRV致病机制等相关研究奠定了基础。 展开更多
关键词 伪狂犬病病毒 小鼠 背根神经节 转录组测序 差异转录基因
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SIRT6对肝细胞脂质合成的影响及其转录基因分析
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作者 曹海超 张松 +3 位作者 丁美玲 马澜婧 刘巧云 聂勇战 《现代生物医学进展》 CAS 2016年第15期2831-2835,共5页
目的:在肝脏L-02 SIRT6-KO细胞系中,观察SIRT6在肝脏脂滴形成中的作用,初步研究在肝脏脂质代谢中SIRT6相关的转录差异基因发挥作用的分子机制。方法:利用基因敲除技术TALEN技术,构建肝脏L-02 SIRT6特异性敲除细胞系,并经q PCR和Western ... 目的:在肝脏L-02 SIRT6-KO细胞系中,观察SIRT6在肝脏脂滴形成中的作用,初步研究在肝脏脂质代谢中SIRT6相关的转录差异基因发挥作用的分子机制。方法:利用基因敲除技术TALEN技术,构建肝脏L-02 SIRT6特异性敲除细胞系,并经q PCR和Western blot检测其表达水平;利用油酸刺激L-02 SIRT6-KO细胞及其对照组,体外模拟肝脏脂肪变的条件,并经高内涵和油红染色验证其脂滴形成能力;利用基因芯片检测L-02 SIRT6-KO及其对照细胞系中相关差异基因,并经生物信息学分析筛选脂代谢相关差异基因。结果:建立人肝脏SIRT6特异性敲除细胞系,q PCR显示m RNA下降50%,但Western blot证明SIRT6完全敲除;BODIPY实验及油红O染色发现,L-02 SIRT6-KO细胞比对照组其脂滴形成数量增多,高内涵荧光检测显示L-02 SIRT6-KO细胞中脂滴荧光强度比对照组增强(176.38±2.55 vs 104.26±2.08);通过对差异转录基因分析,发现了一组在脂质代谢中和SIRT6相关的关键基因如STEAP4、HMGA2、PDE1A、INSL4等。结论:成功建立人肝脏L-02 SIRT6-KO细胞系,并经实验证明SIRT6缺失能够促进脂滴形成;筛选到一组和SIRT6相关参与脂质代谢的关键基因。 展开更多
关键词 TALEN L-02 SIRT6-KO 脂质代谢 差异转录基因
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非洲猪瘟病毒MGF110-5-6L基因功能分析 被引量:1
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作者 张柯慧 葛海亮 +3 位作者 于少雄 李连峰 李素 仇华吉 《中国兽医学报》 CAS CSCD 北大核心 2023年第7期1357-1365,共9页
非洲猪瘟(African swine fever, ASF)是由非洲猪瘟病毒(African swine fever virus, ASFV)感染猪只引起的一种高度接触性传染病,最急性型致死率可高达100%。ASFV结构非常复杂,编码超过160多种蛋白质,其中多基因家族(multigene families,... 非洲猪瘟(African swine fever, ASF)是由非洲猪瘟病毒(African swine fever virus, ASFV)感染猪只引起的一种高度接触性传染病,最急性型致死率可高达100%。ASFV结构非常复杂,编码超过160多种蛋白质,其中多基因家族(multigene families, MGFs)蛋白包括MGF100、MGF110、MGF300、MGF360和MGF505/530等,且MGF360和MGF505可抑制宿主干扰素(interferon, IFN)的产生以及影响ASFV的致病性。但关于MGF110家族相关基因的功能尚无报道,因此本研究选取了MGF110家族中MGF110-5-6L基因进行了相关功能的探究。应用同源重组技术构建了MGF110-5-6L基因缺失的突变体病毒(ASFV-ΔMGF110-5-6L),通过检测该突变体病毒在原代猪肺泡巨噬细胞(porcine alveolar macrophages, PAMs)中的复制水平来探究其在ASFV复制周期中的功能,结果表明,其复制动力学曲线与亲本病毒ASFV HLJ/2018(ASFV-WT)一致,说明缺失MGF110-5-6L基因不影响ASFV在PAMs中的复制。为了探究MGF110-5-6L蛋白在ASFV复制周期中的生物学功能,将ASFV-WT和ASFV-ΔMGF110-5-6L分别感染PAMs,在感染后4,12,20 h收集样品,进行转录组测序。结果显示,与ASFV-WT感染组相比,ASFV-ΔMGF110-5-6L感染组中差异表达基因主要为和IL-17信号通路、肿瘤坏死因子(TNF)信号通路以及细胞因子与细胞因子受体之间的相互作用相关。本研究构建了ASFV MGF110-5-6L基因缺失毒株,并通过转录组测序发现,MGF110-5-6L基因作为ASFV复制非必需基因具有拮抗细胞因子表达的功能,不仅为探究MGF110家族成员的生物学功能提供科学数据,还为深入揭示ASFV拮抗宿主细胞天然免疫反应的分子机制提供参考。 展开更多
关键词 非洲猪瘟病毒 MGF110-5-6L 转录组测序 差异转录基因
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5种单核苷酸多态性与厦门地区汉族人群类风湿关节炎易感性的关系 被引量:4
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作者 徐振兴 陈进春 +1 位作者 邱明山 滕菁 《广东医学》 CAS 北大核心 2016年第7期1003-1006,共4页
目的探讨5种单核苷酸多态性(SNP)与汉族人群类风湿关节炎(RA)易感性的关系。方法收集91例RA患者和100例健康人血液标本,利用基因测序法检测rs2230926、rs7574865、rs3761847、rs2228145和rs2234067等5种SNPs,同时用ELISA法检测RA患... 目的探讨5种单核苷酸多态性(SNP)与汉族人群类风湿关节炎(RA)易感性的关系。方法收集91例RA患者和100例健康人血液标本,利用基因测序法检测rs2230926、rs7574865、rs3761847、rs2228145和rs2234067等5种SNPs,同时用ELISA法检测RA患者的血清抗CCP。分为RA组和对照组进行病例对照研究。结果 (1)5种SNP在对照组和RA组中的分布均符合Hardy-Weinberg平衡(P〉0.05)。rs3761847和rs2228145的不同等位基因频率与RA的易感性显著相关[OR(95%CI)分别为1.61(1.07-2.41)、0.64(0.42-0.97),P〈0.05],rs7574865、rs2234067及rs2230926与RA的易感性未见显著相关(P〉0.05)。(2)rs2228145与RA患者血清抗CCP反应性显著相关(P=0.014 9),其C等位基因频率在两组中的差异有统计学意义(P〈0.05);而rs2228145及rs3761847与RA患者血清RF反应性、rs3761847与RA患者血清抗CCP反应性均无显著相关(P〉0.05)。结论厦门地区汉族人rs3761847和rs2228145基因多态性可能与RA的易感性存在关联,而rs7574865、rs2234067及rs2230926则与RA的易感性可能无关。 展开更多
关键词 类风湿关节炎 单核苷酸多态性 肿瘤坏死因子诱导蛋白3 信号传导及转录激活因子4 肿瘤坏死因子受体相关因子1/补体5 白细胞介素-6受体 转录因子Ets差异基因7
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基于转录组分析大肠杆菌响应亚碲酸盐的机制 被引量:1
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作者 胡苏姝 彭万里 +2 位作者 林双君 邓子新 梁如冰 《微生物学报》 CAS CSCD 北大核心 2022年第7期2702-2718,共17页
亚碲酸盐对绝大多数微生物有高毒性,可用作抗菌剂;但其具体毒性机制仍不清楚。【目的】理解亚碲酸盐的毒性机制,揭示亚碲酸盐处理导致的代谢变化。【方法】本研究通过比较转录组分析与挖掘差异转录基因,探讨了大肠杆菌响应亚碲酸盐胁迫... 亚碲酸盐对绝大多数微生物有高毒性,可用作抗菌剂;但其具体毒性机制仍不清楚。【目的】理解亚碲酸盐的毒性机制,揭示亚碲酸盐处理导致的代谢变化。【方法】本研究通过比较转录组分析与挖掘差异转录基因,探讨了大肠杆菌响应亚碲酸盐胁迫的机制。【结果】Escherichia coli MG1655在10μg/mL亚碲酸盐处理1 h后,比较和分析了亚碲酸盐处理组与对照组的转录水平差异,发现细胞呈现一种明显的适应性变化,许多参与重要代谢途径的基因转录水平改变。其中,与核糖体代谢和鞭毛组装相关基因的转录水平发生显著变化,表明这两条途径很可能是亚碲酸盐作用的主要途径。与细胞能动性、金属离子代谢、细胞膜功能相关的基因的转录水平也发生了明显变化,可能是由于其参与了抵抗亚碲酸盐毒性的细胞代谢调节和损伤修复。【结论】本项工作有助于推动亚碲酸盐毒性机理的研究,促进亚碲酸盐的临床应用。 展开更多
关键词 亚碲酸盐 比较转录 差异转录基因 核糖体代谢 鞭毛组装
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De novo assembly and comparative analysis of root transcriptomes from different varieties of Panax ginseng C. A. Meyer grown in different environments 被引量:6
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作者 ZHEN Gang ZHANG Lei +7 位作者 DU YaNan YU RenBo LIU XinMin CAO FangRui CHANG Qi DENG Xing Wang XIA Mian HE Hang 《Science China(Life Sciences)》 SCIE CAS CSCD 2015年第11期1099-1110,共12页
Panax ginseng C. A. Meyer is an important traditional herb in eastern Asia. It contains ginsenosides, which are primary bioactive compounds with medicinal properties. Although ginseng has been cultivated since at leas... Panax ginseng C. A. Meyer is an important traditional herb in eastern Asia. It contains ginsenosides, which are primary bioactive compounds with medicinal properties. Although ginseng has been cultivated since at least the Ming dynasty to increase production, cultivated ginseng has lower quantities of ginsenosides and lower disease resistance than ginseng grown under natural conditions. We extracted root RNA from six varieties of fifth-year P. ginseng cultivars representing four different growth conditions, and performed Illumina paired-end sequencing. In total, 163,165,706 raw reads were obtained and used to generate a de novo transcriptome that consisted of 151,763 contigs(76,336 unigenes), of which 100,648 contigs(66.3%) were successfully annotated. Differential expression analysis revealed that most differentially expressed genes(DEGs) were upregulated(246 out of 258, 95.3%) in ginseng grown under natural conditions compared with that grown under artificial conditions. These DEGs were enriched in gene ontology(GO) terms including response to stimuli and localization. In particular, some key ginsenoside biosynthesis-related genes, including HMG-Co A synthase(HMGS), mevalonate kinase(MVK), and squalene epoxidase(SE), were upregulated in wild-grown ginseng. Moreover, a high proportion of disease resistance-related genes were upregulated in wild-grown ginseng. This study is the first transcriptome analysis to compare wild-grown and cultivated ginseng, and identifies genes that may produce higher ginsenoside content and better disease resistance in the wild; these genes may have the potential to improve cultivated ginseng grown in artificial environments. 展开更多
关键词 Panax ginseng de novo assembly paired-end sequencing comparative transcriptome analysis ginsenoside biosynthesis disease resistance genes
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Transcriptome profile of human neuroblastoma cells in the hypomagnetic field 被引量:8
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作者 MO WeiChuan LIU Ying +1 位作者 BARTLETT Perry F HE RongQiao 《Science China(Life Sciences)》 SCIE CAS 2014年第4期448-461,1-3,共14页
Research has shown that the hypomagnetic field(HMF)can affect embryo development,cell proliferation,learning and memory,and in vitro tubulin assembly.In the present study,we aimed to elucidate the molecular mechanism ... Research has shown that the hypomagnetic field(HMF)can affect embryo development,cell proliferation,learning and memory,and in vitro tubulin assembly.In the present study,we aimed to elucidate the molecular mechanism by which the HMF exerts its effect,by comparing the transcriptome profiles of human neuroblastoma cells exposed to either the HMF or the geomagnetic field.A total of 2464 differentially expressed genes(DEGs)were identified,216 of which were up-regulated and2248 of which were down-regulated after exposure to the HMF.These DEGs were found to be significantly clustered into several key processes,namely macromolecule localization,protein transport,RNA processing,and brain function.Seventeen DEGs were verified by real-time quantitative PCR,and the expression levels of nine of these DEGs were measured every 6 h.Most notably,MAPK1 and CRY2,showed significant up-and down-regulation,respectively,during the first 6 h of HMF exposure,which suggests involvement of the MAPK pathway and cryptochrome in the early bio-HMF response.Our results provide insights into the molecular mechanisms underlying the observed biological effects of the HMF. 展开更多
关键词 hypomagnetic field geomagnetic field transcriptome profile massively parallel sequencing MAPK1 CRY2
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Transcriptome analysis of blood stasis syndrome in subjects with hypertension 被引量:7
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作者 He Ling Fang Meixia +6 位作者 Chen Liguo Zhou Jianhua Yuan Jing Xu Jing Shan Yan Xu Qingyun Xiong Tingting 《Journal of Traditional Chinese Medicine》 SCIE CAS CSCD 2016年第2期173-180,共8页
OBJECTIVE:To screen for m RNAs associated with blood stasis syndrome and to explore the genetic mechanisms of blood stasis syndrome in hypertension.METHODS:This study involved groups of patients with hypertension and ... OBJECTIVE:To screen for m RNAs associated with blood stasis syndrome and to explore the genetic mechanisms of blood stasis syndrome in hypertension.METHODS:This study involved groups of patients with hypertension and blood stasis,including those with Qi deficiency,Qi stagnation,cold retention and heat retention;as well as hypertensive patients without blood stasis and healthy individuals.Human umbilical vein endothelial cells were co-cultured with the sera of these healthy individuals and patients with blood stasis syndrome.Total RNA was extracted from these cells and assessed by a high-throughput sequencing method(Solexa)and digital gene expression.Differentially expressed genes among these six groups were compared using whole genome sequences,and m RNAs associated with blood stasis syndrome identified.Differences in gene use and gene ontology function were an-alyzed.Genes enriched significantly and their pathways were determined,as were network interactions,and encoded proteins.Gene identities were confirmed by real-time polymerase chain reactions.RESULTS:Compared with cells cultured in sera of the blood stasis groups,those culture in sera of healthy individuals and of the non-blood stasis group showed 11 and 301 differences,respectively in stasis-related genes.Genes identified as differing between the blood stasis and healthy groups included activating transcription factor 4,activating transcription factor 3,DNA-damage inducible transcription factor 3,Tribbles homolog 3,CCAAT/enhancer binding protein-β,and Jun proto-oncogene(JUN).Pathway and protein interaction network analyses showed that these genes were associated with endoplasmic reticulum stress.Cells cultured in sera of patients with blood stasis and Qi deficiency,Qi stagnation,heat retention,and cold retention were compared with cells cultured in sera of patients with the other types blood stasis syndrome.The comparison showed differences in expression of 28,28,34,and 32 specific genes,respectively.CONCLUSION:The pathogenesis of blood stasis syndrome in hypertension is related to endoplasmic reticulum stress and involves the differential expression of the activating transcription factor 4,activating transcription factor 3,DNA-damage inducible transcription factor 3,Tribbles homolog 3,CCAAT/enhancer binding protein-β,and JUN genes. 展开更多
关键词 HYPERTENSION Blood stasis RNA mes senger Endoplasmic reticulum stress
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Transcriptome changes in Polygonum multiflorum Thunb. roots induced by methyl jasmonate
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作者 Hong-chang LIU Wei WU +2 位作者 Kai HOU Jun-wen CHEN Zhi ZHAO 《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》 SCIE CAS CSCD 2015年第12期1027-1041,共15页
Transcriptome profiling has been widely used to analyze transcdptomic variation in plants subjected to abiotic or biotic stresses. Although gene expression changes induced by methyl jasmonate (MeJA) have been profil... Transcriptome profiling has been widely used to analyze transcdptomic variation in plants subjected to abiotic or biotic stresses. Although gene expression changes induced by methyl jasmonate (MeJA) have been profiled in several plant species, no information is available on the MeJA-triggered transcriptome response of Polygonum multiflorum Thunb., a species with highly valuable medicinal properties. In this study, we used transcriptome profiling to investigate transcdptome changes in roots of P. mu/tiflorum seedlings subjected to a 0.25 mmol/L-MeJA rootirrigation treatment. A total of 18 677 differentially expressed genes (DEGs) were induced by MeJA treatment, of which 4535 were up-regulated and 14 142 were down-ragulated compared with controls. These DEGs were associated with 125 metabolic pathways. In addition to various common primary and secondary metabolic pathways, several sec- ondary metabolic pathways related to components with significant pharmacological effects were enriched by MeJA, including arachidonic acid metabolism, linoleic acid metabolism, and stilbenoid biosynthesis. The MeJA-induced transcdptome changes uncovered in this study provide a solid foundation for future study of functional genes controlling effective components in secondary metabolic pathways of P. multiflorum. 展开更多
关键词 Polygonum multiflorum Thunb. Methyl jasmonate Transcriptome change Differentially expressed genes
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