利用DNS法快速分析及优化柚皮苷的酶解过程

【目的】建立采用3,5-二硝基水杨酸(DNS)法快速测定柚皮苷水解率的方法,并利用该方法对柚苷酶催化水解柚皮苷生成柚皮素的反应过程进行优化研究。【方法】利用棘孢曲霉JMUdb058发酵得到的柚苷酶催化水解柚皮苷,采用DNS法对柚皮苷酶解过程中还原糖的生成量进行分析,经过换算得到柚皮苷的水解率,并在此基础上通过单因素实验优化柚皮苷的酶解过程。【结果】在柚皮苷的水解过程中,还原糖的生成量与柚皮苷的水解量及柚皮素的生成量均呈现出良好的线性关系,因此可利用DNS法测定体系中还原糖的生成量,并通过换算得到柚皮素的生成量。利用该方法优化柚皮苷的酶解过程得到柚苷酶转化柚皮苷的最适温度为50°C、pH为5.0、酶用量为8 U/mL、底物浓度为0.2 g/100 mL。在此条件下,柚皮苷酶解150 min后可达到平衡,此时其水解率为85%。通过Lineweaver-Burk双倒数作图法测得Km为     

DDD: Distributed Dataset DNS

Cluster Computing - The advent of inexpensive data storage has resulted in larger and larger datasets as the cost of pruning data becomes more expensive then storing it for future insights. This...     

DNS/DNS-Homologiestudien innerhalb der Gattung Pediococcus

Zusammenfassung An 17 Stämmen der Gattung Pediococcus, die auf Grund charakteristischer physiologisch-biochemischer Merkmale aus einer Anzahl von etwa 500 Kulturen unterschiedlicher Herkunft ausgewählt wurden, werden mit Hilfe der DNS/DNS-Hybridisierung die verwandtschaftlichen Beziehungen untersucht. Dabei wird die Eigenständigeit der Arten P. pentosaceus, P. acidilactici, P. parvulus, P. damnosus und P. halophilus bestätigt. Der für P. acidilactici vorgeschlagene Neotyp-Stamm NCDO 1859 ist jedoch nach den vorliegenden Ergebnissen ein echter P. pentosaceus. Ein Stamm von P. dextrinicus, der früher bereits als P. cerevisiae var. dextrinicus beschrieben wurde, zeigt zu keiner untersuchten Art eine genetische Verwandtschaft auf der Basis der DNS/DNS-Homologie. Zwei aus Bier und Bierhefe isolierte Stämme, die hinsichtlich des Phänotyps P. parvulus nahestehen, zeigen weder zu dieser Art noch zu allen anderen bisher beschriebenen Arten hinsichtlich des Genotyps einen hohen Verwandtschaftsgrad.

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DNA-DNA homology studies on the genus Pediococcus

17 strains of Pediococcus, including new isolates and strains from various collections, were investigated by nucleic acid hybridisation. Most of the newly isolated strains, selected from about 500 strains according to physiological and biochemical characters, could be attributed to known species. The reassociation data demonstrate the separate status of P. pentosaceus, P. acidilactici, P. damnosus, P. parvulus and P. halophilus. The proposed neotype strain of P. acidilactici NCDO 1859, however, has been found to be in reality a genuine P. pentosaceus. P. dextrinicus, formerly described as P. cerevisiae var. dextrinicus showed a remarkable lack of relationship to any of the strains tested. Two isolates exhibiting a high phenotypic similarity to P. parvulus show little genetic relationship to this and especially to all other species.

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Abkürzungen CTAB N-cetyl-N,N,N-trimethyammoniumbromid - EDTA Äthylendiamintetraessigsäure - PPO 2,5-Diphenyloxazol - POPOP 1,4-Bis-(5-phenyl-2-oxazolyl)-benzol - SSC Saline-Trinatriumcitrat (0,15M NaCl, 0,015M Trinatriumcitrat, pH 7,0) - Tris Tris(hydroxymethyl)-aminomethan - T _m mittlere Schmelztemperatur der DNS Adresse für Sonderdruck-Anforderungen     

血浆游离氨基酸与支/芳比值DNS荧光法测定

<正> 近年来,随着各种氨基酸代谢疾病早期诊断的需要,血浆支链与芳香族氨基酸浓度比值(BCAA/AAA)作为慢性肝病诊断指标的提出,以及氨酸输液疗法的开展,临床上迫切需要对血浆及其它体液的游离氨基酸进行定量分析。先进的自动分析仪器价格昂贵,经典的纸层析—茆三酮法又难以选到要求,因而目前在普通医院中尚难开展此项定量工作。     

Heterochromatin,repetitive DNS und Nukleolen in Leberzellen von Microtus agrestis

Zusammenfassung Die Zellstruktur von Leberzellen der Erdmaus, Microtus agrestis, wurde nach Giemsafärbung, Feulgenbehandlung, Behandlung mit Ribonuklease und nach Färbung des konstitutiven Heterochromatins untersucht. Das konstitutive Heterochromatin ist in Leberzellen nicht heteropyknotisch, das fakultative Heterochromatin ist im weiblichen Geschlecht als Sexchromatinkörperchen sichtbar. Bestimmungen des relativen DNS-Gehalts ergaben, daß die Zahl der Sexchromatinkörperchen der Ploidie der Zellkerne proportional ist. Die Nukleolen liegen in Hepatozyten oft randständig; in 59% der diploiden Zellkerne sind 2 Nukleolen enthalten. Nach Anfärbung der repetitiven DNS werden oft auch die Nukleolen gefärbt, nach Ribonukleasebehandlung tritt dieser Effekt nicht auf. Das konstitutive Heterochromatin wird in Form von 2 langen fädigen Strukturen sichtbar.

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Heterochromatin, repetitive DNA and nucleoli in liver cells of Microtus agrestis

Summary The nuclear structure of parenchymal liver cells of embryo and adult Microtus agrestis was studied in smear and section preparations after staining with Giemsa solution and treatment according to Feulgen, after treatment with ribonuclease and after specific staining of constitutive heterochromatin. In liver cell nuclei only the facultative heterochromatin is heteropycnotic, a sex chromatin body is observable in female but not in male animals. Constitutive heterochromatin is not heteropycnotic in liver cells. Measurements of the relative DNA content showed that nuclei with one sex chromatin body are diploid; tetraploid nuclei possess two and octoploid nuclei four sex chromatin bodies. Solely in the diploid cell nuclei of the intrahepatic gall ducts two large chromocenters are found. The nucleoli in hepatocytes often lie at the perimeter of the nucleus. 17% of the diploid nuclei contain one nucleolus, 59% two nucleoli, 23% three and 1% four. After staining of repetitive DNA, the nucleoli often become stained as well; after treatment with ribonuclease this effect does not occur. The constitutive heterochromatin becomes visible in form of two long, threadlike structures. After longer periods of dissociation the sex chromatin body ceases to be visible. Sex chromatin and constitutive heterochromatin are contiguous to the nucleoli.

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Mit dankenswerter Unterstützung durch das Bundesministerium für Bildung und Wissenschaft der Bundesrepublik Deutschland.     

Verteilung und Aufteilung der DNS bei einigen Cyanophyceen,festgestellt durch ihre DAPI-Fluoreszenz

The DNA of Blue-Green Algae may be specifically and most efficiently elucidated by the combined use of the fluorochrome DAPI and DNAse treatment. InChroococcus turgidus andSynechococcus aeruginosus it forms a network which usually is positioned at the periphery of the centroplasm. At the beginning of cell division it seems to be invaginated by the ingrowing cell wall. Later it must be distributed by other means because two equal parts are connected by a number of fine and straight DNA threads. InOscillatoria limosa usually all parts of the centroplasm are interspersed with the DNA network. In surface view, the DNA occasionally seems to consist of a number of independant rods, but in reality these are connected to the general network. Division apparantly occurs by ingrowth of the crosswall and usually results in equal daughter nuclear equivalents although occasionally they are unequal. Four strains ofMicrocoleus vaginatus were found to have nuclear equivalents of the same general appearance as inOscillatoria limosa.

     

研究荧光标记BCAA及AAA在大鼠体内代谢

使用DNS-Cl标记BCAA及AAA等六种氨基酸,观察这些氨基酸在大鼠小肠的吸收,血液的清除以及肝脏、肌肉、肾脏、脑等器官的摄取及排空情况。结果:小肠在30分钟开始吸收,4小时部分排空;血液在4小时开始清除,16小时清除完毕;肝脏、肌肉、肾脏、脑组织等,都在30分钟全部或部分摄取,但各器官对不同氨基酸,排空的时间不尽相同。     

Eine Methode zur histochemischen Bestimmung von RNS und DNS an der gleichen Zelle

Zusammenfassung Es wird eine Methode beschrieben, die die quantitative cytophotometrische Bestimmung von DNS und RNS nebeneinander am gleichen Objekt gestattet. Dazu werden die Zellen mit Gallocyaninchromalaun und anschließend mit der Feulgen-Reaktion gefärbt, wobei ein Peulgenfarbstoff verwendet wird, der im blauen Spektralbereich absorbiert. Auch unter den veränderten Färbebedingungen verlaufen beide Parbreaktionen quantitativ. Es werden für die Durchführung der Methode die geeigneten Versuchsansätze und ein Verfahren zur Auswertung der cytophotometrischen Meßergebnisse angegeben.

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Microphotometric determination of DNA and RNA within the same cell

Summary A method is described which allows the simultanous cytophotometric determination of DNA and RNA within the same cell. Cells are stained with gallocyanine chromealum and then, in an additional step, by a Feulgen procedure in which Coriphosphin 0 was used as a Schiff type reagent to accomplish absorption in the blue region of the spectrum. It could be shown that under the altered conditions both staining reactions can be used for quantitative purposes. Methods for carrying out the preparation of the material, for cytophotometric measurements and for calculating RNA and DNA values are suggested.

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Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.

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Wir widmen diese Arbeit dem Andenken Wolfgang Zellers. Wesentliche Teile dieser Arbeit waren als Teil seiner Dissertation vorgesehen. Sein tragischer Tod hat seine Promotion verhindert.     

阿特拉津降解菌株DNS32的降解特性及分类鉴定与降解途径研究

【目的】研究阿特拉津降解菌株DNS32的菌种分类、降解特性及降解途径,丰富阿特拉津降解菌菌种资源。【方法】在长期施用阿特拉津的东北地区寒地黑土中筛选出一株以阿特拉津为唯一氮源生长的降解菌株DNS32,测定其基本降解特性,通过16S rRNA序列分析进行分类鉴定,并利用阿特拉津降解基因PCR扩增技术及降解产物生成量的测定,进一步揭示其降解途径。【结果】实验结果发现DNS32菌株具有较好的降解能力,且在相对较低温度下也具有一定的降解能力。16S rRNA序列分析结果表明DNS32与鲁氏不动杆菌(Acinetobacter lwoffii)16S rRNA序列同源性高达99%。成功地扩增降解基因trzN、atzB及atzC,实验结果表明DNS32遵循Arthrobacter aurescens TC1的降解模式,可将阿特拉津降解为氰尿酸,降解产物的生成量测定也证明了这一点。【结论】实验结果丰富了阿特拉津降解菌菌种资源,为不动杆菌属的阿特拉津降解菌研究提供了参考。     

Trennung der effekte von transmutation und strahlung nach einbau von radionukliden in die DNS

Summary Among the various methods for studying the relative effects of transmutation and radiation of incorporated nuclides, simulation of beta radiation by external gamma exposure is of practical importance.Self-irradiation and mutual irradiation of the labeled cells cannot be neglected in any case. Furthermore, additional hypothetical and experimental problems may arise from using either external beta radiation or different isotopes of an element.By means of external gamma irradiation on the other hand, this being equivalent to the internal beta radiation from a microdosimetrical point of view, the radiation effect of the nuclide alone can be observed without any modification of other experimental parameters. To determine such equivalent gamma radiation for labeled cell nuclei ofVicia faba roots, the authors applied the Monte Carlo Method to the beta spectra of³²P,³H,¹⁴C and¹³¹J, to the energy-dependent LET and to different cell diameters. The existence of secondary particle equilibrium inside the nuclei during gamma exposure was assumed. For certain radionuclides and cell sizes it is possible to calculate gamma spectra which induce energy spectra in the nuclei similar to those caused by the beta particles originating in the nuclear DNA.

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Trennung der effekte von transmutation und strahlung nach einbau von radionukliden in die DNS

Zusammenfassung Unter den verschiedenen Methoden zur Untersuchung der relativen Wirkung von Transmutation und Strahlung inkorporierter Nuklide kommt der Simulation der Betastrahlung durch eine externe Gammastrahlung praktische Bedeutung zu. Nicht immer kann man nämlich davon ausgehen, daß die Selbstbestrahlung der zu untersuchenden Objekte vernachlässigt werden kann. Auch bringt sowohl die Verwendung einer äußeren Betastrahlung als die verschiedener Isotope desselben Elements häufig zusätzliche prinzipielle und experimentelle Probleme mit sich. Hingegen kann durch eine der Betastrahlung äquivalente Gammastrahlung der Strahleneffekt des Nuklids allein ohne Veränderung der Kultur- und Inkorporationsbedingungen untersucht werden. Allerdings ist es keineswegs einfach, diese äquivalente Gammastrahlung auf rein theoretischem Wege zu ermitteln. Immerhin können mit Hilfe des skizzierten Verfahrens zumindest in einigen Fällen Gammaspektren berechnet werden, die im Innern des Zellkerns vonVicia faba, Energieabsorptionsspektren erzeugen, die denen von im Zellkern emittierten Betateilchen ähnlich sind.Unter Berücksichtigung des in der Diskussion Gesagten läßt sich das Verfahren sicherlich verbessern. Auch fßr andere Objekte und Objektverteilungen dürften sich dann äußere Strahlenquellen ermitteln lassen, mit denen der relative Anteil von Transmutation und Strahlung an der Gesamtwirkung des radioaktiven Zerfalls untersucht werden kann.

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Eingegangen am 26. August 1974     

阿特拉津降解菌Acinetobacter sp. DNS32对无机氮源的响应

【目的】研究Acinetobacter sp.DNS32的生长、阿特拉津降解能力和降解基因转录水平的表达对无机氮素的响应关系,为菌株的工程应用提供指导与理论基础。【方法】以Acinetobactersp.DNS32为对象,采用摇瓶法研究菌株在阿特拉津培养基中菌株生长情况及降解能力对外加硝态氮与铵态氮的响应关系,利用荧光定量PCR技术检测DNS32降解基因表达量对外加无机氮源的响应关系。【结果】外加无机氮源可以促进DNS32菌株的生长,提高阿特拉津降解能力,无机氮源对DNS32菌株的trzN、atzB和atzC 3种降解基因表达均有促进作用,加入无机氮源的试验处理中DNS32菌株trzN基因的表达量最高可达对照的11.252±2.408倍,推断DNS32菌株的这3种降解基因所编码的酶是稳定表达的组成酶。【结论】DNS32降解阿特拉津不受"氮饥饿"诱导机制调控,且无机氮源的存在对菌株的生长与降解有促进作用,因此菌株在土壤修复实践中具有广阔的应用前景。     

Eine Methode zum Nachweis von hydrolytischen Enzymen mit sukzedaner zytophotometrischer Bestimmung von DNS,Histon und Gesamtprotein

Zusammenfassung Bei der zytophotometrischen Bestimmung von Kernproteinen an gemischten Zellpopulationen wie Lymphknoten, Knochenmark oder Milz ist die Zuordnung der jeweils zu untersuchenden Einzelzelle zu einer bestimmten Zellrasse oft schwierig. Durch diesen Unsicherheitsfaktor kann die Aussagekraft derartiger Messungen stark eingeschränkt werden. Um diese Fehlermöglichkeit zu vermindern, wird ein Verfahren beschrieben, welches durch Anwendung zytochemischer Enzymnachweismethoden eine zuverlässige Klassifizierung der zu messenden Einzelzelle vor der anschließend vorzunehmenden eigentlichen quantitativen Analyse derselben Zelle erlaubt, ohne die Meßergebnisse zu beeinträchtigen.

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A method for the demonstration of hydrolytic enzymes with sequential cytophotometric measurement of DNA, nuclear histone and total protein

Summary In cytophotometric analyses of nuclear proteins of mixed cell populations such as lymph nodes, bone marrow or spleen it is frequently difficult to place a single cell to be explored into a precise category of cell types. This factor of uncertainty concerning the classification of cells may substantially reduce the value of those measurements. In order to eliminate or diminish such sources of error a method is described which by means of cytochemical enzyme tracing allows for a reliable classification of single cells to be examined prior to the real quantitative analysis of the same cell without impairment of the results of the measurements.

     

Über die Adsorption von DNS in der Grenzfläche Quecksilber–Elektrolyt

The adsorption of deoxyribonucleic acid (DNA) in the mercury–electrolyte interface has been investigated. The effect of this adsorption on the differential capacity of the electrical double layer between a polarized mercury surface and an 0.15M NaCl solution containing DNA was measured by means of the alternating current polarography (Breyer polarography). The effective alternating current ? under actual conditions (adsorption processes only, small electrolytic resistance, small alternating current frequency, and alternating current amplitude) is directly proportional to the differential double layer capacity. The combination of this method with the application of a stationary mercury drop electrode allows the coverage of the electrode to be followed, continuously in the range 0.2 sec, to about 60 sec. The diffusion is the rate-controlled step of the adsorption kinetics. Therefore the lowering of the alternating current ? by the adsorbed DNA is proportional to the surface concentration for partly covered surface and reaches a constant value after the surface becomes fully covered. Adsorption of further layers does not affect the differential capacity. This makes it possible to determine the maximum surface concentration of the DNA. For that it is necessary to determine the diffusion coefficient of DNA. This was done directly by Strassburger and Reinert in our institute. The surface concentrations of the native DNA and the relative surface concentrations of the denatured DNA in dependence on the potential of the polarized mercury surface was estimated. Both surface concentrations show a pronounced dependence on the potential with a minimum of the surface concentration around ?0.4 V with respect to the normal calomel electrode. This property may be caused by the structure of the adsorption layer depending on the potential. That means that only several segments at the rigid DNA molecules are adsorbed and the other ones remain in the solution near the surface. The adsorption in the neighborhood of the electrocapillary zero potential at ?0.4 V is strongest, and therefore the fraction of the adsorbed segments has a maximum. At these potentials consequently the maximum coverage is already reached at relatively low surface concentrations. Opposite to this is Miller's hypothesis, that native DNA preserves its double helical structure when adsorbed on a negatively charged mercury surface, whereas unfolding occurs on a positively charged mercury surface. Miller's hypothesis is supported by facts that the surface concentration of the denatured DNA should be independent of the potential and should be equal to the surface concentration of the native DNA at a positively charged mercury surface. But an evaluation of Miller's diagrams by no means gives an independence on the potential of the surface concentration of the denatured DNA and no accordance between the surface concentrations of denatured and of native DNA's at the positively charged mercury surface. Moreover Miller compared different DNA samples with different moleculer weights and possibly with different molecular weight distributions. Both the molecular weight and the molecular weight distribution have a pronounced influence on the surface concentration. Therefore this accordance mentioned above is not evident. The critical inspection of Miller's work and the own investigation lead to the conclusion that an unfolding or denaturation of native DNA does not take place in the mercury–electrolyte interface.     

Exclusion of the dysplastic nevus syndrome (DNS) locus from the short arm of chromosome 1 by linkage studies in Dutch families

Familial dysplastic nevus syndrome (DNS) is an autosomal dominant premalignant condition characterized by multiple large moles of variable size and color and a strongly increased risk for cutaneous malignant melanoma. In order to determine the chromosomal localization of the DNS gene, linkage studies were initiated in six large Dutch families. No support was obtained for linkage between the loci for DNS and the rhesus blood group on chromosome 1. Data from additional markers (DNF15S1, D1Z2, FUCA1, D1S17, D1S57, and PGM1) make it possible to exclude the DNS gene from the short arm of chromosome 1 in these Dutch families.     

Metabolic ability and gene characteristics of Arthrobacter sp. strain DNS10, the sole atrazine-degrading strain in a consortium isolated from black soil

Strain DNS10 was the only member that could utilize atrazine as the sole nitrogen source for growth in an atrazine-degrading consortium which was isolated from black soil previously in our laboratory. It belongs to the genus Arthrobacter according to the sequence of 16S rRNA gene and is designated as Arthrobacter sp. DNS10. 16S rRNA gene phylogenetic analysis showed that strain DNS10 was located in a different evolutionary branch comparing with other Arthrobacter sp. atrazine-degrading strains. The degrading genes such as trzN, atzB and atzC harbored in strain DNS10 revealed high sequence similarity with those in Arthrobacter aurescens TC1 and Pseudomonas sp. ADP. These genes enabled the strain DNS10 to decompose atrazine to cyanuric acid. This was further proved by the results that the strain DNS10 (10⁸ CFU mL⁻¹) could degrade the whole atrazine (100 mg L⁻¹) in the medium within 24 h at 30 °C and there was 66.13 ± 2.11 mg L⁻¹ cyanuric acid accumulated at 24 h. These results imply that the strain DNS10 seems to be an excellent atrazine-degrading strain. Furthermore, this paper helps us in the better understanding of the strain evolution by comparing the metabolic ability and gene characteristics of strain DNS10 with other geographically distinct atrazine-degrading strains.