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1.
Umami is one of the five basic tastes and an important taste attribute of green tea.Amino acids are major components contributing to umami taste and the top three amino acids in tea are theanine,glutarnic acid and aspartic acid.Trace of umami taste components adenosine monophosphate (AMP) and guanosine monophosphate (GMP) were detected in tea.Levels of umami taste components varied with tea cultivar,environmental conditions,fertilizer and processing method.The taste threshold of amino acids differentiated greatly and its DOT (dose over threshold) is an important indicator evaluating its contribution to umami taste.  相似文献   

2.
农杆菌介导glgC-TM基因转化水稻研究   总被引:6,自引:1,他引:6  
 利用农杆菌介导将glgC-TM基因导入多个水稻品种。研究表明农杆菌转化过程中,潮霉素是一种很好的筛选剂,成熟胚以及国产的羧苄青霉素不适宜用作农杆菌介导的遗传转化。不同品种以及不同的质粒在转化过程中对抗性愈伤的得率和阳性植株的转化率不存在显著影响。T1代PCR检测表明,在转基因当代就有可能获得转基因纯系。  相似文献   

3.
EGCG与锌离子互作对PC-3细胞能量代谢的影响   总被引:1,自引:0,他引:1  
采用Hoechst33258荧光染色法观察EGCG、Zn2+和EGCG+Zn2+对前列腺癌PC-3细胞凋亡的诱导作用;采用HPLC法检测了EGCG、Zn2+和EGCG+Zn2+对PC-3细胞内腺苷酸(ATP、ADP、AMP)含量的影响,并分析了能量负荷(EC)和ATP/ADP比值。结果表明,Hoechst33258染色后正常细胞发出均匀微弱的蓝色荧光,细胞核未见异常,EGCG、Zn2+和EGCG+Zn2+处理后凋亡的PC-3细胞都发出较强的蓝色荧光,细胞核DNA断裂及染色体高度浓缩;EGCG、Zn2+及二者混合物处理后细胞内总腺苷酸含量、EC及ATP/ADP比值都下降,表明EGCG与Zn2+对PC-3细胞能量代谢都存在明显的抑制作用。  相似文献   

4.
胆固醇对水稻幼苗抗冷性的影响   总被引:2,自引:0,他引:2  
低浓度的胆固醇处理水稻种子,可增强冷害水稻幼苗的超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性,降低丙二醛(MDA)含量,提高可溶性蛋白总量中的热稳定蛋白含量,但过氧化物酶(POD)活性变化不明显。亚细胞组分测定表明,ATP主要存在于叶绿体和线粒体中,胆固醇对低温下叶绿体的保护作用尤为明显。  相似文献   

5.
 以水稻为材料,系统探讨了水稻中(脱氧)胞苷脱氨酶基因的组织表达特性和胁迫诱导表达特性。实时PCR结果表明,7个编码(脱氧)胞苷脱氨酶的基因在苗期水稻根、茎和叶中的表达具有组织特异性,但这些基因在水稻茎中的表达都低于根和叶中的表达;同时,这些基因胁迫诱导表达特性也不尽相同,只有Os07g0245100的表达同时受到低温、干旱、高盐和病害侵染等多种胁迫的诱导。因此,(脱氧)胞苷酸脱氨酶在水稻中可能通过调节体内嘧啶核苷的水平从而在水稻的环境胁迫应答反应中发挥重要调控作用。  相似文献   

6.
胶乳再生过程中腺苷水平(ATP、ADP和AMP含量)能够反映橡胶树乳管的生理代谢强度。运用高效液相色谱法,成功建立了一种简单、快速的橡胶树胶乳中ATP、ADP和AMP的定量方法,并采用该方法对胶乳再生中的腺苷水平进行分析。结果表明:胶乳再生过程中,早期(0~12 h)乳管主要利用排胶后剩余的AMP和ADP合成ATP,此后先后恢复AMP和ADP(6 h后)以及ATP(12 h后)的合成能力,并在排胶后12~48 h之间出现一个明显的ATP快速合成期。对排胶影响面的能荷分析表明,排胶造成割线下方2~7 cm区  相似文献   

7.
孙化鹏  钟晓红  乔飞 《热带作物学报》2018,39(11):2200-2206
甲羟戊酸焦磷酸脱羧酶(Mevalonate diphosphate decarboxylase,MVD)是三萜皂苷生物合成途径中的关键酶, 为研究其在洋常春藤中的基因功能,通过 RT-PCR 和 RACE 技术从洋常春藤叶片中克隆获得 HhMVD 基因的 cDNA 全 长序列,并通过 RT-qPCR 技术分析其表达规律。结果表明:RACE 克隆获得的 HhMVD 基因 cDNA 序列全长 1 799 bp, 包含一个完整开放阅读框(ORF)1 263 bp、5′非编码区(5′UTR)192 bp、3′非编码区(3′UTR)344 bp;该基因编码 420 个氨基酸,分子质量 46.6 ku,理论等电点为 6.57,不含跨膜区,属于非分泌型蛋白;HhMVD 蛋白具有 GHMP 激 酶 N 端结构域,属于甲羟戊酸激酶系列,与刺五加、人参、三七等同科植物亲缘关系较近。RT-qPCR 结果表明,洋常 春藤中 HhMVD 基因的时空表达相对稳定,但与常春藤皂苷含量差异变化之间存在一定的负相关性。洋常春藤 HhMVD 基因的成功克隆及表达分析研究,为深入探讨其基因功能、阐明其在常春藤皂苷生物合成途径中的作用提供理论依据。  相似文献   

8.
采用3′RACE和RT-PCR技术克隆芒果核苷二磷酸激酶基因全长c DNA序列,并用实时荧光定量PCR分析该基因的表达模式。结果表明:Mi NDPK1基因c DNA全长1 019 bp,开放阅读框为447 bp,编码148个氨基酸。Mi NDPK1蛋白定位于细胞质,无信号肽和跨膜区域,具有典型的核苷二磷酸激酶活性结构域和其他磷酸化活性位点;与麻疯树、橡胶树同源性最高为89%,与菠菜同源性最低为82%。Mi NDPK1在芒果各组织中均有表达,以叶片中表达最高,其次是花和果皮;在芒果进入生殖时期的花芽分化期表达量最高,此后在芒果果实发育进程中逐渐降低并趋于平稳。结合病害处理转录组样本中的差异性表达结果,推测芒果Mi NDPK1基因在花芽分化进程、抗病性诱导及免疫应答通路上发挥重要作用。  相似文献   

9.
采用RT-PCR和RACE(Rapid amplification of cDNA ends)技术,从艾纳香(Blumea balsamifera L.DC)的叶片中克隆到单萜化合物合成的关键酶牻牛儿基焦磷酸合成酶(BbGPPS)基因。研究结果显示,BbGPPS基因的cDNA全长1 692 bp,包含开放阅读框(ORF)1 083 bp,编码361个氨基酸;亚细胞结构定位于叶绿体,既非膜蛋白也非分泌性蛋白。疏水性分析显示,BbGPPS是亲水性蛋白。同源性比对结果显示,BbGPPS蛋白与其他植物中GPPS蛋白具有高度的相似性,且具有异戊烯基结构域。系统发育分析表明,所有序列被聚为5大类,BbGPPS与其他菊科植物聚类一类,与万寿菊(Tagetes erecta)TeGPPS亲缘关系最近,其次是甜菊(Stevia rebaudiana)SrGPPS。  相似文献   

10.
以薄层色谱法对光周期不同处理的马铃薯不同时间的叶片,匍匐茎以及刚形成的小块茎中6-氨基嘌呤和腺嘌呤核苷的变化进行了分析,探讨了该二种代谢产物对块茎形成的调控作用.发现其含量以某种规律变化,可以促进块茎形成.  相似文献   

11.
牻牛儿基牻牛儿基焦磷酸合酶(Geranylgeranyl diphosphate synthase,GGPS)是萜类合成途径的结构酶,对植物生长发育具有重要意义。本研究通过RACE和RT-PCR方法克隆得到5条潜在的茶树GGPS序列,分别命名为CsGGPS1-4和CsGGPS9,其中CsGGPS9存在3条等位基因,分别是CsGGPS9-1、CsGGPS9-2和CsGGPS9-3,在系统进化树上与其他基因分成两支。蛋白质序列分析表明,茶树GGPS家族成员都具有polyprenyl_synt结构域,不存在信号肽序列。亚细胞定位预测结果显示,CsGGPS1、CsGGPS2和CsGGPS4定位在叶绿体上,CsGGPS3和CsGGPS9定位在线粒体上。通过Swiss Model进行三维建模,结合"three-floor"模型对茶树GGPS家族成员的功能进行预测,预测结果显示,CsGGPS1、CsGGPS2和CsGGPS4是GGPS;CsGGPS3是异源二聚体形式的牻牛儿基焦磷酸合酶的小亚基;CsGGPS9的催化主产物是碳链数大于30的异戊烯基焦磷酸。q RT-PCR分析表明,CsGGPS1整体表达丰度较低,仅在一芽二叶中表达量稍高;CsGGPS2在茶树各个组织中均有表达,在花中表达量最高,且花发育过程中表达量先上升后下降;CsGGPS3在叶和幼根中的表达量高于花,花发育过程中表达平稳;CsGGPS4在茶树各个组织中表达量数值相近,在花发育过程中表达量变化趋势与CsGGPS2相同;CsGGPS9的表达量在成熟叶中显著低于幼嫩叶片。  相似文献   

12.
13.
目的:建立测定抱茎苦荬菜药材中总黄酮和腺苷的方法。方法:采用UV—VIS法测定总黄酮的含量。采用RP—HPLC法测定腺苷的含量,色谱条件为:XDB—C18(150mm×4.6mm,5μm)色谱柱,以乙腈-水(体积比为6:94)为流动相,检测波长为260nm,流速为1ml/min^-1。结果:腺苷进样量在0.072—3.6μg范围内线性关系良好(r=0.999),平均回收率为100.3%,相对标准差(RSD)为1.35%。结论:方法准确、可靠,适合测定抱茎苦荬菜药材中总黄酮和腺苷的含量。  相似文献   

14.
CMP-Neu5Ac is an important sugar nucleotide for biosynthesis of sialic acid and its conjugates. In this paper, a large-scale production system of CMP-Neu5Ac by a single strain is reported. The co-expression of Neu5Ac aldolase (EC4.1.3.3) and CMP-Neu5Ac synthetase (EC 2.7.7.43) was achieved by constructing individual genes into one plasmid and having a single culture that has both NeuAc aldolase and CMP-Neu5Ac synthetase activities. Overall this system only employed N-acetylmannosamine, excess of pyruvate and CTP to produce CMP-Neu5Ac. This work has demonstrated that a large-scale synthesis of sialic acid-derived oligosaccharides could be achieved economically and efficiently through a single, biosynthetic pathway engineered microorganism.  相似文献   

15.
This study was designed to investigate the effects and underlying mechanisms of Astaxanthin (AST) on high-fructose-induced hyperuricemia (HUA) from the perspectives of the uric acid (UA) synthesis and excretion in rat models. Following six weeks of a 10% fructose diet, the level of serum UA effectively decreased in the AST groups as compared to the model group. The enzymatic activities of xanthine oxidase (XOD) and adenosine deaminase (ADA) were significantly inhibited, and the mRNA expression levels of XOD and ADA significantly decreased after the AST administration. These results suggested that the AST reduced UA synthesis by inhibiting the mRNA expressions and enzyme activities of XOD and ADA, thereby contributing to HUA improvement. On the hand, the relative expressions of the mRNA and protein of kidney reabsorption transport proteins (GLUT9 and URAT1) were significantly down-regulated by AST, while that of the kidney secretion proteins (OAT1, OAT3 and ABCG2) were significantly up-regulated by AST. These results indicated that the AST promoted UA excretion by regulating the urate transport proteins, and thus alleviated HUA. This study suggested that the AST could serve as an effective alternative to traditional medicinal drugs for the prevention of fructose-induced HUA.  相似文献   

16.
硫是植物生长所必需的大量元素之一。植物通过根系吸收土壤中的硫酸盐,经ATP硫酸化酶活化为腺苷5'-磷酰硫酸(adenosine 5'-phosphosulfate,APS)。APS的代谢包括初级代谢和次级代谢,分别是APS还原酶催化APS生成亚硫酸盐;腺苷5'-磷酰硫酸激酶(adenosine 5'-phosphosulfate kinase,APSK)催化APS磷酸化生成3'-磷酸-腺苷5'-磷酰硫酸(3'-phosphoadenosine 5'-phosphosulfate,PAPS),PAPS进一步作为硫酸根供体参与胞内的硫酸化反应。近年来的研究表明,拟南芥APSK在调节硫的初级和次级代谢的过程中具有重要功能,且受氧化胁迫的APSK1在亚基C86和C119间形成二硫键降低其催化活性。水稻的同工酶是否也会受到氧化还原调控尚无相关报道。对APSK序列进行分析,发现水稻APSK1含有与拟南芥APSK1 C86和C69同源的半胱氨酸。本研究对水稻APSK1基因进行了克隆、双突变和酶活分析。OsAPSK1及双突变C36A/C69A经原核表达、纯化,获得了OsAPSK1及双突变蛋白。体外酶活分析表明,在氧化环境中OsAPSK1活性受到抑制,而C36A/C69A双突变蛋白不受氧化环境的影响。推测水稻胞内受到氧化胁迫时会降低OsAPSK1的活性,从而促进还原型谷胱甘肽的合成,提高水稻的抗氧化能力。通过分析PAPS的含量与氧化胁迫之间的相关性及不同OsAPSK亚型的活性调节机制等将有利于进一步阐明APSK在胞内的功能。  相似文献   

17.
Sulfate can be activated by ATP sulfurylase and adenosine 5'-phosphosulfate kinase(APSK) in vivo. Recent studies suggested that APSK in Arabidopsis thaliana regulated the partition between APS reduction and phosphorylation and its activity can be modulated by cellular redox status. In order to study regulation of APSK in rice(Os APSK), Os APSK1 gene was cloned and its activity was analyzed. Os APSK1 C36 and C69 were found to be the conserved counterparts of C86 and C119, which involved in disulfide formation in At APSK. C36A/C69 A Os APSK1 double mutation was made by site directed mutagenesis. Os APSK1 and its mutant were prokaryotically over-expressed and purified, and then assayed for APS phosphorylation activity. Os APSK1 activity was depressed by oxidized glutathione, while the activity of its mutant was not. Further studies in the case that oxidative stress will fluctuate in vivo 3'-phosphoadenosine-5'-phosphosulfate content, and all APSK isoenzymes have similar regulation patterns are necessary to be performed.  相似文献   

18.
Extracellular ATP mediates proinflammatory and antiproliferative effects via activation of P2 nucleotide receptors. In contrast, its metabolite, the nucleoside adenosine, is strongly immunosuppressive and enhances tumor proliferation and metastasis. The conversion of ATP to adenosine is catalyzed by ectonucleotidases, which are expressed on immune cells and typically upregulated on tumor cells. In the present study, we identified sulfopolysaccharides from brown and red sea algae to act as potent dual inhibitors of the main ATP-hydrolyzing ectoenzymes, ectonucleotide pyrophosphatase/phosphodiesterase-1 (NPP1) and ecto-nucleoside triphosphate diphosphohydrolase-1 (NTPDase1, CD39), showing nano- to picomolar potency and displaying a non-competitive mechanism of inhibition. We showed that one of the sulfopolysaccharides tested as a representative example reduced adenosine formation at the surface of the human glioblastoma cell line U87 in a concentration-dependent manner. These natural products represent the most potent inhibitors of extracellular ATP hydrolysis known to date and have potential as novel therapeutics for the immunotherapy of cancer.  相似文献   

19.
In spite of the high abundance and species diversity of diatoms, only a few bioactive compounds from them have been described. The present study reveals a high number of mammalian cell death inducing substances in biofilm-associated diatoms sampled from the intertidal zone. Extracts from the genera Melosira, Amphora, Phaeodactylum and Nitzschia were all found to induce leukemia cell death, with either classical apoptotic or autophagic features. Several extracts also contained inhibitors of thrombin-induced blood platelet activation. Some of this activity was caused by a high content of adenosine in the diatoms, ranging from 0.07 to 0.31 μg/mg dry weight. However, most of the bioactivity was adenosine deaminase-resistant. An adenosine deaminase-resistant active fraction from one of the extracts was partially purified and shown to induce apoptosis with a distinct phenotype. The results show that benthic diatoms typically found in the intertidal zone may represent a richer source of interesting bioactive compounds than hitherto recognized.  相似文献   

20.
Studies have indicated the presence of both a de novo biosynthesis of new polyisoprene chains and the elongation of pre-existing rubber-PP chains in rubber particles from Ficus elastica Roxb. and small rubber particles from Hevea brasiliensis (Willd. ex A. Juss.) Müll. Arg. In this paper, we have isolated rubber particles from the cortical parenchyma of Parthenium argentatum Gray with an adsorbed isopentenyl pyrophosphate isomerase that was competent to form polyisoprene to determine the presence of de novo biosynthesis of polyisoprene and the elongation of pre-existing rubber-PP. The gel filtrated rubber particles were supplied saturating concentrations of [14C]-isopentenyl pyrophosphate for 1 h and the resulting concentrations of [14C]-polyisoprene were isolated and analyzed with a Waters 150C Gel Permeation Chromatograph. The mol wt distribution of the [14C]-polyisoprene ranged from 103 to 106 Da with no formation of high mol wt chains equal to the polyisoprene in the mol wt distribution of natural rubber from P. argentatum. The mol wt distribution of [14C]-polyisoprene following 1 h of incubation of the reaction mixture was similar to the mol wt distribution formed by the bound rubber transferase in washed rubber particles of H. brasiliensis and the time-dependent formation of polyisoprene by washed rubber particles from P. argentatum. The data support the conclusion that the washed rubber particles from P. argentatum contain a bound rubber transferase that initiates the formation of new chains of polyisoprene from the dimethylallyl pyrophosphate initiator generated from the added isopentenyl pyrophosphate isomerase followed by the extensive polymerization of isopentenyl pyrophosphate to form a monomodal rubber polymer. There was no evidence in these washed rubber particles of a chain elongation of pre-existing rubber-PP.  相似文献   

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