TY - THES A1 - Wettstein, Christoph T1 - Cytochrome c-DNA and cytochrome c-enzyme interactions for the construction of analytical signal chains N2 - Electron transfer (ET) reactions play a crucial role in the metabolic pathways of all organisms. In biotechnological approaches, the redox properties of the protein cytochrome c (cyt c), which acts as an electron shuttle in the respiratory chain, was utilized to engineer ET chains on electrode surfaces. With the help of the biopolymer DNA, the redox protein assembles into electro active multilayer (ML) systems, providing a biocompatible matrix for the entrapment of proteins. In this study the characteristics of the cyt c and DNA interaction were defined on the molecular level for the first time and the binding sites of DNA on cyt c were identified. Persistent cyt c/DNA complexes were formed in solution under the assembly conditions of ML architectures, i.e. pH 5.0 and low ionic strength. At pH 7.0, no agglomerates were formed, permitting the characterization of the NMR spectroscopy. Using transverse relaxation-optimized spectroscopy (TROSY)-heteronuclear single quantum coherence (HSQC) experiments, DNAs’ binding sites on the protein were identified. In particular, negatively charged AA residues, which are known interaction sites in cyt c/protein binding were identified as the main contact points of cyt c and DNA. Moreover, the sophisticated task of arranging proteins on electrode surfaces to create functional ET chains was addressed. Therefore, two different enzyme types, the flavin dependent fructose dehydrogenase (FDH) and the pyrroloquinoline quinone dependent glucose dehydrogenase (PQQ-GDH), were tested as reaction partners of freely diffusing cyt c and cyt c immobilized on electrodes in mono- and MLs. The characterisation of the ET processes was performed by means of electrochemistry and the protein deposition was monitored by microgravimetric measurements. FDH and PQQ-GDH were found to be generally suitable for combination with the cyt c/DNA ML system, since both enzymes interact with cyt c in solution and in the immobilized state. The immobilization of FDH and cyt c was achieved with the enzyme on top of a cyt c monolayer electrode without the help of a polyelectrolyte. Combining FDH with the cyt c/DNA ML system did not succeed, yet. However, the basic conditions for this protein-protein interaction were defined. PQQ-GDH was successfully coupled with the ML system, demonstrating that that the cyt c/DNA ML system provides a suitable interface for enzymes and that the creation of signal chains, based on the idea of co-immobilized proteins is feasible. Future work may be directed to the investigation of cyt c/DNA interaction under the precise conditions of ML assembly. Therefore, solid state NMR or X-ray crystallography may be required. Based on the results of this study, the combination of FDH with the ML system should be addressed. Moreover, alternative types of enzymes may be tested as catalytic component of the ML assembly, aiming on the development of innovative biosensor applications. N2 - In den Energiegewinnungsprozessen der Zellen spielen biochemische Reaktion, die auf Elektronentransfer (ET) basieren, eine wichtige Rolle. So sind die Proteinkomplexe der Atmungskette, welche an der inneren Membran der Mitochondrien abläuft, über eine ET-Kette miteinander verbunden. In biotechnologischen Anwendungen wird dieses Phänomen genutzt um Proteine auf der Oberfläche von Elektroden als funktionierende ET-Ketten zu arrangieren. Dabei kann der ET innerhalb dieser Kaskaden als elektrischer Strom gemessen und als Signal betrachtet werden. Dies ermöglicht die Anwendung von proteinmodifizierten Elektroden als Biosensoren und Biobrennstoffzellen. Ein geeigneter Baustein für den Aufbau vielschichtiger ET-Systeme ist das kleine, eisenhaltige Protein Cytochrom c (Cyt c), welches in der Lage ist Elektronen aufzunehmen, zu transportieren und wieder abzugeben. Als zweiter Baustein dient das lange, fadenartige Biomolekül DNA. DNA und Cyt c interagieren unter bestimmten Bedingungen aufgrund ihrer entgegengesetzten Oberflächenladungen. Dies ermöglicht den schichtweisen Aufbau stabiler Cyt c/DNA-Multischichten (MS) auf Elektrodenoberflächen, welche durch die sogenannte Layer-by-Layer (LbL) Technik aufgebaut werden. In diesen MS Systemen behält Cyt c trotz der Immobilisierung seine Beweglichkeit um die eigene Achse, wodurch der Selbstaustausch von Elektronen zwischen den Cyt c Molekülen sowie der ET zur Elektrode gewährleistet wird. Der molekulare Aufbau der Cyt c/DNA MS sowie die Interaktion zwischen den zwei biologischen Bausteine ist weitgehend unerforscht, daher wurden in der vorliegenden Studie die genauen Bedingungen der Cyt c/DNA Interaktion in Lösung untersucht. Außerdem wird die Eignung des MS Systems zur Einbettung von Enzymen getestet. Die Bausteine des MS-Systems, Cyt c und DNA bilden in Lösung stabile Komplexe unter den Assemblierungsbedingungen der MS (d.h. pH 5.0 und geringe Salzkonzentration). Im Vergleich dazu tritt bei pH 7.0 eine schwächere Interaktion auf, die für eine Komplexbildung nicht ausreicht. Dies ermöglicht die Untersuchung der Interaktion mittels Kernspinresonanzspektroskopie (NMR, engl. nuclear magnetic resonance spectroscopy), wobei die Interaktionsstellen des DNA-Moleküls auf Cyt c bestimmt werden. Im Vergleich zu pH 7.0 wird im leicht sauren pH-Bereich (6.0) eine erhöhte Anzahl an Interaktionspunkten gefunden, was Rückschlüsse auf eine erhöhte Interaktion zulässt. Dies resultiert schließlich in der starken Bindung bei pH 5.0, die den Aufbau stabiler Cyt c/DNA-MS auf Elektrodenoberflächen ermöglicht. Darüber hinaus spielen der Salzgehalt der Lösung sowie das Konzentrationsverhältnis von Cyt c und DNA eine wichtige Rolle. Auf der Grundlage des Cyt c/DNA-MS Aufbaus sollte durch die Kopplung eines Enzymes eine Signalkette mit sensorischen Eigenschaften geschaffen werden. Das Enzym dient dabei als Erkennungselement für bestimmte Moleküle in Lösung. Durch die Reaktion des Enzyms mit dem Molekül wird ein bioelektrisches Signal generiert, das durch elektrochemische Methoden gemessen wird. Dies wurde mit zwei verschiedenen Enzymen, der Glukose Dehydrogenase (GDH) und der Fruktose Dehydrogenase (FDH), untersucht. Beide Enzyme waren in der Lage mit einer Cyt c Monoschicht zu kommunizieren und konnten mit dem redox Protein auf der Elektrodenoberfläche immobilisiert werden. GDH konnte erfolgreich mit dem Cyt c/DNA-MS System gekoppelt und die Sensoreigenschaften der so aufgebauten Elektronentransferkette charakterisiert werden. Zusammenfassend charakterisiert diese Arbeit die Bedingungen der Cyt c/DNA-Komplexbildung und gibt einen Einblick in die bisher unbekannte Interaktion zwischen Cyt c und DNA auf der molekularen Ebene. Darüber hinaus wird die Nutzbarkeit des Cyt c/DNA MS Systems zur Einbettung von Enzymen am Beispiel der GDH gezeigt und schafft somit die Grundlage für das bessere Verständnis von ET Reaktionen zwischen Proteinen auf Elektrodenoberflächen. T2 - Cytochrom c-DNA und Cytochrom c-Enzym Interaktion für den Aufbau analytischer Signalketten KW - biosensor KW - protein KW - DNA KW - enzyme KW - interaction KW - DNA KW - Biosensor KW - Enzym KW - Interaktion KW - Protein Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-78367 ER - TY - THES A1 - Vogel, Stefanie T1 - Sequence dependency of photon and electron induced DNA strand breaks T1 - Sequenzabhängigkeit von photonen-und elektroneninduzierten DNA Strangbrüchen N2 - Deoxyribonucleic acid (DNA) is the carrier of human genetic information and is exposed to environmental influences such as the ultraviolet (UV) fraction of sunlight every day. The photostability of the DNA against UV light is astonishing. Even if the DNA bases have a strong absorption maximum at around 260 nm/4.77 eV, their quantum yield of photoproducts remains very low 1. If the photon energies exceed the ionization energy (IE) of the nucleobases ( ̴ 8-9 eV) 2, the DNA can be severely damaged. Photoexcitation and -ionization reactions occur, which can induce strand breaks in the DNA. The efficiency of the excitation and ionization induced strand breaks in the target DNA sequences are represented by cross sections. If Si as a substrate material is used in the VUV irradiation experiments, secondary electrons with an energy below 3.6 eV are generated from the substrate. This low energy electrons (LEE) are known to induce dissociative electron attachment (DEA) in DNA and with it DNA strand breakage very efficiently. LEEs play an important role in cancer radiation therapy, since they are generated secondarily along the radiation track of ionizing radiation. In the framework of this thesis, different single stranded DNA sequences were irradiated with 8.44 eV vacuum UV (VUV) light and cross sections for single strand breaks (SSB) were determined. Several sequences were also exposed to secondary LEEs, which additionally contributed to the SSBs. First, the cross sections for SSBs depending on the type of nucleobases were determined. Both types of DNA sequences, mono-nucleobase and mixed sequences showed very similar results upon VUV radiation. The additional influence of secondarily generated LEEs resulted in contrast in a clear trend for the SSB cross sections. In this, the polythymine sequence had the highest cross section for SSBs, which can be explained by strong anionic resonances in this energy range. Furthermore, SSB cross sections were determined as a function of sequence length. This resulted in an increase in the strand breaks to the same extent as the increase in the geometrical cross section. The longest DNA sequence (20 nucleotides) investigated in this series, however, showed smaller cross section values for SSBs, which can be explained by conformational changes in the DNA. Moreover, several DNA sequences that included the radiosensitizers 5-Bromouracil (5BrU) and 8-Bromoadenine (8BrA) were investigated and the corresponding SSB cross sections were determined. It was shown that 5BrU reacts very strongly to VUV radiation leading to high strand break yields, which showed in turn a strong sequence-dependency. 8BrA, on the other hand, showed no sensitization to the applied VUV radiation, since almost no increase in strand breakage yield was observed in comparison to non-modified DNA sequences. In order to be able to identify the mechanisms of radiation damage by photons, the IEs of certain DNA sequences were further explored using photoionization tandem mass spectrometry. By varying the DNA sequence, both the IEs depending on the type of nucleobase as well as on the DNA strand length could be identified and correlated to the SSB cross sections. The influence of the IE on the photoinduced reaction in the brominated DNA sequences could be excluded. N2 - Desoxyribonukleinsäure (DNA) ist als Träger der menschlichen Erbinformation täglich vielen Einflüssen ausgesetzt. Diese Einflüsse können Teil unserer Umwelt sein, wie der ultraviolette (UV) Anteil des Sonnenlichts. Die Photostabilität der DNA gegen UV-Licht ist erstaunlich, denn trotz eines starkes Absorptionsmaximum der DNA-Basen bei etwa 260 nm/4,77 eV, bleibt ihre Quantenausbeute an Photoprodukten sehr gering 1. Überschreiten die Photonenenergien die Ionisationsenergie (IE) der Nukleinbasen ( ̴ 8-9 eV) 2, kann die DNA schwer geschädigt werden. Es treten Anregungs- und Ionisierungsreaktionen auf, die zu Strangbrüchen in der DNA führen. Die Effizienz der induzierten Strangbrüche in den untersuchten DNA-Sequenzen wird durch Wirkungsquerschnitte dargestellt. Wird in den Bestrahlungsexperimenten Silizium als Substratmaterial verwendet, werden aus dem Substrat zusätzliche Sekundärelektronen mit einer Energie unter 3,6 eV erzeugt, die weiteren Schaden an der DNA verursachen. Diese niederenergetischen Elektronen (LEE) sind dafür bekannt, dissoziative Elektronenanlagerung (DEA) und damit Strangbrüche in der DNA zu erzeugen. LEEs entstehen sekundär entlang des Strahlungsweges von ionisierender Strahlung im biologischen Gewebe, wenn in der Behandlung der Krankheit Krebs Strahlentherapie eingesetzt wird. Im Rahmen dieser Arbeit wurden verschiedene Einzelstrang-DNA-Sequenzen mit 8.44 eV Vakuum-UV (VUV) Licht bestrahlt und Wirkungsquerschnitte für Einzel-strangbrüche (SSB) bestimmt. Ein Teil der Sequenzen wurde außerdem sekundär erzeugten LEEs ausgesetzt, die einen zusätzlichen Beitrag zu den SSBs liefern. Als erstes wurde der Wirkungsquerschnitt für SSBs in Abhängigkeit der Nukleinbasen bestimmt. Hierbei weisen sowohl die DNA Sequenzen, die nur ein Sorte an Nukleinbasen besitzen als auch die gemischte Sequenzen sehr ähnliche Werte auf. Durch den zusätzlichen Einfluss der LEEs hat sich wiederum für die DNA Sequenzen mit nur einer Sorte an Nukleinbasen ein stark ausgeprägter Trend gezeigt. Die Polythymin-Sequenz weist den höchsten Wirkungsquerschnitt für SSBs auf, was durch ausgeprägte anionische Resonanzen in diesem Energiebereich begründet werden kann. Des Weiteren wurden Wirkungsquerschnitte für SSBs in Abhängigkeit Sequenzlänge ermittelt. Dabei ergab sich eine Erhöhung der SSBs im gleichen Maße wie die Vergrößerung des geometrischen Wirkungsquerschnitts. Die längste DNA Sequenz (20 Nukleotide), die in dieser Reihe untersucht wurde, zeigte hingegen kleinere Werte für den SSB Wirkungsquerschnitt, was durch Konformationsänderungen in der DNA erklärt werden kann. Einige der untersuchten DNA Sequenzen wurden zusätzlich mit den Radiosensibilisatoren 5-Bromouracil (5BrU) und 8-Bromoadenine (8BrA) modifiziert und entsprechende SSB Wirkungsquerschnitte bestimmt. Hierbei hat sich gezeigt, dass 5BrU mittels einer hohen Strangbruchausbeute sehr stark auf VUV Strahlung reagiert, wobei das Ausmaß der Reaktion stark sequenzabhängig ist. 8BrA hingegen, weist keine Sensibilisierung gegenüber der verwendeten VUV Strahlung auf, da keine Erhöhung der Strangbruchausbeute gegenüber unmodifizierten DNA Sequenzen ersichtlich ist. Um die Mechanismen der Strahlenschädigung durch Photonen besser einschätzen zu können, wurden zusätzlich die IEs bestimmter DNA Sequenzen mit Hilfe der Photoionisations-Tandem-Massenspektrometrie untersucht. Durch Variation der DNA-Sequenzen konnte sowohl ein Trend der IEs in Abhängigkeit der Nukleinbasen und der DNA-Stranglänge identifiziert und als auch eine Abhängigkeit der Reaktivität von 5BrU von seinem IE in der entsprechenden DNA Sequenz ausgeschlossen werden. Die IE Trends und die Wirkungsquerschnitte für SSBs wurden abschließend in Korrelation gebracht. KW - DNA KW - photo ionization KW - dissociative electron attachment KW - DNA origami KW - radiosensitizer KW - ionization energy KW - tandem mass spectrometry KW - DNS KW - Photoionisation KW - Dissoziative Elektronenanlagerung KW - DNA Origami KW - Radiosensibilisator KW - Ionisierungsenergie KW - Tandemmassenspektrometrie Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-419669 ER - TY - THES A1 - Möser, Christin T1 - Modular DNA constructs for oligovalent bio-enhancement and functional screening T1 - Modulare DNA-Konstrukte für oligovalente Bio-Verstärkung und funktionelles Screening N2 - Deoxyribonucleic acid (DNA) nanostructures enable the attachment of functional molecules to nearly any unique location on their underlying structure. Due to their single-base-pair structural resolution, several ligands can be spatially arranged and closely controlled according to the geometry of their desired target, resulting in optimized binding and/or signaling interactions. This dissertation covers three main projects. All of them use variations of functionalized DNA nanostructures that act as platform for oligovalent presentation of ligands. The purpose of this work was to evaluate the ability of DNA nanostructures to precisely display different types of functional molecules and to consequently enhance their efficacy according to the concept of multivalency. Moreover, functionalized DNA structures were examined for their suitability in functional screening assays. The developed DNA-based compound ligands were used to target structures in different biological systems. One part of this dissertation attempted to bind pathogens with small modified DNA nanostructures. Pathogens like viruses and bacteria are known for their multivalent attachment to host cells membranes. By blocking their receptors for recognition and/or fusion with their targeted host in an oligovalent manner, the objective was to impede their ability to adhere to and invade cells. For influenza A, only enhanced binding of oligovalent peptide-DNA constructs compared to the monovalent peptide could be observed, whereas in the case of respiratory syncytial virus (RSV), binding as well as blocking of the target receptors led to an increased inhibition of infection in vitro. In the final part, the ability of chimeric DNA-peptide constructs to bind to and activate signaling receptors on the surface of cells was investigated. Specific binding of DNA trimers, conjugated with up to three peptides, to EphA2 receptor expressing cells was evaluated in flow cytometry experiments. Subsequently, their ability to activate these receptors via phosphorylation was assessed. EphA2 phosphorylation was significantly increased by DNA trimers carrying three peptides compared to monovalent peptide. As a result of activation, cells underwent characteristic morphological changes, where they "round up" and retract their periphery. The results obtained in this work comprehensively prove the capability of DNA nanostructures to serve as stable, biocompatible, controllable platforms for the oligovalent presentation of functional ligands. Functionalized DNA nanostructures were used to enhance biological effects and as tool for functional screening of bio-activity. This work demonstrates that modified DNA structures have the potential to improve drug development and to unravel the activation of signaling pathways. N2 - Desoxyribonukleinsäure (DNS, engl. DNA) - Nanostrukturen ermöglichen die Anbringung funktioneller Moleküle an nahezu jede einzigartige Stelle der zugrunde liegenden Struktur. Aufgrund der Basenpaar-Strukturauflösung von DNA können mehrere Moleküle (z.B. Liganden) entsprechend der Geometrie ihres gewünschten Ziels räumlich angeordnet und genau kontrolliert werden, was zu optimierten Bindungs- und/oder Signalwechselwirkungen führt. Diese Dissertation umfasst drei Hauptprojekte. Alle Projekte verwenden Varianten von funktionalisierten DNA-Nanostrukturen, die als Plattform für die oligovalente Präsentation von Liganden dienen. Ziel der vorliegenden Arbeit war es, die Fähigkeit von DNA-Nanostrukturen zur präzisen Positionierung verschiedener Arten von funktionellen Molekülen zu evaluieren und folglich die Wirksamkeit der Moleküle gemäß dem Konzept der Multivalenz zu erhöhen. Außerdem wurde untersucht, wie funktionalisierte DNA-Strukturen in verschiedenen Verfahren zur Erforschung von biologischen Interaktionen eingesetzt werden können. Die entwickelten DNA-basierten Liganden wurden verwendet, um Strukturen auf verschiedenen biologischen Systemen gezielt zu binden. In einem Teil dieser Dissertation wurde versucht, Krankheitserreger mit kleinen modifizierten DNA-Nanostrukturen zu binden. Pathogene, wie Viren und Bakterien, sind für ihre multivalente Anheftung an Wirtszellmembranen bekannt. Durch die oligovalente Blockierung ihrer Rezeptoren für die Erkennung und/oder Fusion mit ihrem Wirt sollte ihre Fähigkeit, sich an Zielzellen anzuheften und in diese einzudringen, beeinträchtigt werden. Bei Influenza A Viren konnte nur eine verstärkte Bindung von oligovalenten Peptid-DNA-Konstrukten im Vergleich zu monovalenten Peptiden beobachtet werden, wohingegen bei Respiratorischen Synzytial-Viren (RSV) sowohl die Bindung als auch die Blockierung der Zielrezeptoren zu einer verstärkten Hemmung der Infektion in vitro führte. Im letzten Teil wurden chimäre DNA-Peptidkonstrukte auf ihre Fähigkeit, an Signalrezeptoren auf der Oberfläche von Zellen zu binden und diese zu aktivieren, getestet. Die spezifische Bindung von mit bis zu drei Peptiden konjugierten DNA-Trimeren an EphA2-Rezeptor-exprimierende Zellen wurde in Durchflusszytometrie-Experimenten untersucht. Anschließend wurde ihre Fähigkeit, diese Rezeptoren durch Phosphorylierung zu aktivieren, beurteilt. Die Phosphorylierung von EphA2 war durch DNA-Trimere, die drei Peptide trugen, im Vergleich zu monovalenten Peptiden signifikant erhöht. Infolge der Aktivierung kommt es zu charakteristischen morphologischen Veränderungen der Zellen, bei denen diese ihre Peripherie "abrunden" und zurückziehen. Die in dieser Arbeit erzielten Ergebnisse beweisen umfassend die Fähigkeit von DNA-Nanostrukturen, als stabile, biokompatible, kontrollierbare Plattformen für die oligovalente Präsentation funktioneller Liganden zu fungieren. Funktionalisierte DNA-Nanostrukturen wurden zur Verstärkung biologischer Effekte und als Werkzeug für das funktionelle Screening von biologischen Interaktionen verwendet. Diese Arbeit zeigt, dass modifizierte DNA-Strukturen das Potenzial haben, die Medikamentenentwicklung zu verbessern und die Aktivierung von Signalwegen zu entschlüsseln. KW - DNA KW - multivalency KW - influenza KW - respiratory syncytial virus KW - nanostructure KW - ephrin KW - DNA KW - Ephrin KW - Influenza KW - Multivalenz KW - Nanostruktur KW - Respiratorisches Synzytial-Virus KW - DNS Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-507289 ER - TY - GEN A1 - Meyer, Matthias A1 - Palkopoulou, Eleftheria A1 - Baleka, Sina Isabelle A1 - Stiller, Mathias A1 - Penkman, Kirsty E. H. A1 - Alt, Kurt W. A1 - Ishida, Yasuko A1 - Mania, Dietrich A1 - Mallick, Swapan A1 - Meijer, Tom A1 - Meller, Harald A1 - Nagel, Sarah A1 - Nickel, Birgit A1 - Ostritz, Sven A1 - Rohland, Nadin A1 - Schauer, Karol A1 - Schüler, Tim A1 - Roca, Alfred L. A1 - Reich, David A1 - Shapiro, Beth A1 - Hofreiter, Michael T1 - Palaeogenomes of Eurasian straight-tusked elephants challenge the current view of elephant evolution T2 - Postprints der Universität Potsdam Mathematisch-Naturwissenschaftliche Reihe N2 - The straight-tusked elephants Palaeoloxodon spp. were widespread across Eurasia during the Pleistocene. Phylogenetic reconstructions using morphological traits have grouped them with Asian elephants (Elephas maximus), and many paleontologists place Palaeoloxodon within Elephas. Here, we report the recovery of full mitochondrial genomes from four and partial nuclear genomes from two P. antiquus fossils. These fossils were collected at two sites in Germany, Neumark-Nord and Weimar-Ehringsdorf, and likely date to interglacial periods similar to 120 and similar to 244 thousand years ago, respectively. Unexpectedly, nuclear and mitochondrial DNA analyses suggest that P. antiquus was a close relative of extant African forest elephants (Loxodonta cyclotis). Species previously referred to Palaeoloxodon are thus most parsimoniously explained as having diverged from the lineage of Loxodonta, indicating that Loxodonta has not been constrained to Africa. Our results demonstrate that the current picture of elephant evolution is in need of substantial revision. T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 790 KW - genome sequence KW - woolly mammoth KW - Palaeoloxodon-antiquus KW - phylogenetic analysis KW - African elephants KW - DNA KW - Pleistocene KW - alignment KW - ancient KW - reveal Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-440139 SN - 1866-8372 IS - 790 ER - TY - JOUR A1 - Li, Chenhong A1 - Corrigan, Shannon A1 - Yang, Lei A1 - Straube, Nicolas A1 - Harris, Mark A1 - Hofreiter, Michael A1 - White, William T. A1 - Naylor, Gavin J. P. T1 - DNA capture reveals transoceanic gene flow in endangered river sharks JF - Proceedings of the National Academy of Sciences of the United States of America N2 - For over a hundred years, the "river sharks" of the genus Glyphis were only known from the type specimens of species that had been collected in the 19th century. They were widely considered extinct until populations of Glyphis-like sharks were rediscovered in remote regions of Borneo and Northern Australia at the end of the 20th century. However, the genetic affinities between the newly discovered Glyphis-like populations and the poorly preserved, original museum-type specimens have never been established. Here, we present the first (to our knowledge) fully resolved, complete phylogeny of Glyphis that includes both archival-type specimens and modern material. We used a sensitive DNA hybridization capture method to obtain complete mitochondrial genomes from all of our samples and show that three of the five described river shark species are probably conspecific and widely distributed in Southeast Asia. Furthermore we show that there has been recent gene flow between locations that are separated by large oceanic expanses. Our data strongly suggest marine dispersal in these species, overturning the widely held notion that river sharks are restricted to freshwater. It seems that species in the genus Glyphis are euryhaline with an ecology similar to the bull shark, in which adult individuals live in the ocean while the young grow up in river habitats with reduced predation pressure. Finally, we discovered a previously unidentified species within the genus Glyphis that is deeply divergent from all other lineages, underscoring the current lack of knowledge about the biodiversity and ecology of these mysterious sharks. KW - freshwater sharks KW - DNA KW - museum specimens Y1 - 2015 U6 - https://doi.org/10.1073/pnas.1508735112 SN - 0027-8424 VL - 112 IS - 43 SP - 13302 EP - 13307 PB - National Acad. of Sciences CY - Washington ER - TY - JOUR A1 - Laux, Eva-Maria A1 - Ermilova, Elena A1 - Pannwitz, Daniel A1 - Gibbons, Jessica A1 - Hölzel, Ralph A1 - Bier, Frank Fabian T1 - Dielectric Spectroscopy of Biomolecules up to 110 GHz JF - Frequenz N2 - Radio-frequency fields in the GHz range are increasingly applied in biotechnology and medicine. In order to fully exploit both their potential and their risks detailed information about the dielectric properties of biological material is needed. For this purpose a measuring system is presented that allows the acquisition of complex dielectric spectra over 4 frequency decade up to 110 GHz. Routines for calibration and for data evaluation according to physicochemical interaction models have been developed. The frequency dependent permittivity and dielectric loss of some proteins and nucleic acids, the main classes of biomolecules, and of their sub-units have been determined. Dielectric spectra are presented for the amino acid alanine, the proteins lysozyme and haemoglobin, the nucleotides AMP and ATP, and for the plasmid pET-21, which has been produced by bacterial culture. Characterisation of a variety of biomolecules is envisaged, as is the application to studies on protein structure and function. KW - dielectric KW - spectroscopy KW - permittivity KW - protein KW - DNA KW - amino acid KW - plasmid Y1 - 2018 U6 - https://doi.org/10.1515/freq-2018-0010 SN - 0016-1136 SN - 2191-6349 VL - 72 IS - 3-4 SP - 135 EP - 140 PB - De Gruyter CY - Berlin ER - TY - JOUR A1 - Laux, Eva-Maria A1 - Bier, Frank Fabian A1 - Hölzel, Ralph T1 - Dielectrophoretic Stretching of DNA JF - DNA Nanotechnology N2 - The spatial control of DNA and of self-assembled DNA constructs is a prerequisite for the preparation of DNA-based nanostructures and microstructures and a useful tool for studies on single DNA molecules. Here we describe a protocol for the accumulation of dissolved lambda-DNA molecules between planar microelectrodes by the action of inhomogeneous radiofrequency electric fields. The resulting AC electrokinetic forces stretch the DNA molecules and align them parallel to the electric field. The electrode preparation from off-the-shelf electronic components is explained, and a detailed description of the electronic setup is given. The experimental procedure is controlled in real-time by fluorescence microscopy. KW - Alignment KW - Dielectrophoresis KW - DNA KW - Electrokinetics KW - Interdigitated electrodes KW - Stretching Y1 - 2018 SN - 978-1-4939-8582-1 SN - 978-1-4939-8581-4 U6 - https://doi.org/10.1007/978-1-4939-8582-1_14 SN - 1064-3745 SN - 1940-6029 SP - 199 EP - 208 PB - Humana Press Inc. CY - New York ET - 2 ER - TY - JOUR A1 - Kienzler, Andrea Altevogt Nee A1 - Flehr, Roman A1 - Gehne, Sören A1 - Kumke, Michael Uwe A1 - Bannwarth, Willi T1 - Verification and biophysical characterization of a New Three-Color Forster Resonance-Energy-Transfer (FRET) System in DNA JF - Helvetica chimica acta N2 - We report on a new three-color FRET system consisting of three fluorescent dyes, i.e., of a carbostyril (=quinolin-2(1H)-one)-derived donor D, a (bathophenanthroline)ruthenium complex as a relay chromophore A1, and a Cy dye as A2 (FRET=Forster resonance-energy-transfer) (cf. Fig. 1). With their widely matching spectroscopic properties (cf. Fig. 2), the combination of these dyes yielded excellent FRET efficiencies. Furthermore, fluorescence lifetime measurements revealed that the long fluorescence lifetime of the Ru complex was transferred to the Cy dye offering the possibility to measure the whole system in a time-resolved mode. The FRET system was established on double-stranded DNA (cf. Fig. 3) but it should also be generally applicable to other biomolecules. KW - Forster resonance energy transfer (FRET) system KW - DNA KW - Fluorescence KW - Ruthenium complexes Y1 - 2012 U6 - https://doi.org/10.1002/hlca.201100460 SN - 0018-019X VL - 95 IS - 4 SP - 543 EP - 555 PB - Wiley-VCH CY - Weinheim ER - TY - GEN A1 - Kasyanenko, Nina A1 - Unksov, Ivan A1 - Bakulev, Vladimir A1 - Santer, Svetlana T1 - DNA interaction with head-to-tail associates of cationic surfactants prevents formation of compact particles T2 - Postprints der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe N2 - Cationic azobenzene-containing surfactants are capable of condensing DNA in solution with formation of nanosized particles that can be employed in gene delivery. The ratio of surfactant/DNA concentration and solution ionic strength determines the result of DNA-surfactant interaction: Complexes with a micelle-like surfactant associates on DNA, which induces DNA shrinkage, DNA precipitation or DNA condensation with the emergence of nanosized particles. UV and fluorescence spectroscopy, low gradient viscometry and flow birefringence methods were employed to investigate DNA-surfactant and surfactant-surfactant interaction at different NaCl concentrations, [NaCl]. It was observed that [NaCl] (or the Debye screening radius) determines the surfactant-surfactant interaction in solutions without DNA. Monomers, micelles and non-micellar associates of azobenzene-containing surfactants with head-to-tail orientation of molecules were distinguished due to the features of their absorption spectra. The novel data enabled us to conclude that exactly the type of associates (together with the concentration of components) determines the result of DNA-surfactant interaction. Predomination of head-to-tail associates at 0.01 M < [NaCl] < 0.5 M induces DNA aggregation and in some cases DNA precipitation. High NaCl concentration (higher than 0.8 M) prevents electrostatic attraction of surfactants to DNA phosphates for complex formation. DAPI dye luminescence in solutions with DNA-surfactant complexes shows that surfactant tails overlap the DNA minor groove. The addition of di- and trivalent metal ions before and after the surfactant binding to DNA indicate that the bound surfactant molecules are located on DNA in islets T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 940 KW - azobenzene trimethylammonium bromide KW - head-to-tail surfactant associates KW - DNA KW - ionic strength KW - multivalent ions Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-459806 SN - 1866-8372 IS - 940 ER - TY - JOUR A1 - Kasyanenko, Nina A1 - Unksov, Ivan A1 - Bakulev, Vladimir A1 - Santer, Svetlana T1 - DNA interaction with head-to-tail associates of cationic surfactants prevents formation of compact particles JF - Molecules N2 - Cationic azobenzene-containing surfactants are capable of condensing DNA in solution with formation of nanosized particles that can be employed in gene delivery. The ratio of surfactant/DNA concentration and solution ionic strength determines the result of DNA-surfactant interaction: Complexes with a micelle-like surfactant associates on DNA, which induces DNA shrinkage, DNA precipitation or DNA condensation with the emergence of nanosized particles. UV and fluorescence spectroscopy, low gradient viscometry and flow birefringence methods were employed to investigate DNA-surfactant and surfactant-surfactant interaction at different NaCl concentrations, [NaCl]. It was observed that [NaCl] (or the Debye screening radius) determines the surfactant-surfactant interaction in solutions without DNA. Monomers, micelles and non-micellar associates of azobenzene-containing surfactants with head-to-tail orientation of molecules were distinguished due to the features of their absorption spectra. The novel data enabled us to conclude that exactly the type of associates (together with the concentration of components) determines the result of DNA-surfactant interaction. Predomination of head-to-tail associates at 0.01 M < [NaCl] < 0.5 M induces DNA aggregation and in some cases DNA precipitation. High NaCl concentration (higher than 0.8 M) prevents electrostatic attraction of surfactants to DNA phosphates for complex formation. DAPI dye luminescence in solutions with DNA-surfactant complexes shows that surfactant tails overlap the DNA minor groove. The addition of di- and trivalent metal ions before and after the surfactant binding to DNA indicate that the bound surfactant molecules are located on DNA in islets. KW - azobenzene trimethylammonium bromide KW - head-to-tail surfactant associates KW - DNA KW - ionic strength KW - multivalent ions Y1 - 2018 U6 - https://doi.org/10.3390/molecules23071576 SN - 1420-3049 VL - 23 IS - 7 PB - MDPI CY - Basel ER - TY - JOUR A1 - Heinsohn, Natascha Katharina A1 - Niedl, Robert Raimund A1 - Anielski, Alexander A1 - Lisdat, Fred A1 - Beta, Carsten T1 - Electrophoretic mu PAD for purification and analysis of DNA samples JF - Biosensors : open access journal N2 - In this work, the fabrication and characterization of a simple, inexpensive, and effective microfluidic paper analytic device (mu PAD) for monitoring DNA samples is reported. The glass microfiber-based chip has been fabricated by a new wax-based transfer-printing technique and an electrode printing process. It is capable of moving DNA effectively in a time-dependent fashion. The nucleic acid sample is not damaged by this process and is accumulated in front of the anode, but not directly on the electrode. Thus, further DNA processing is feasible. The system allows the DNA to be purified by separating it from other components in sample mixtures such as proteins. Furthermore, it is demonstrated that DNA can be moved through several layers of the glass fiber material. This proof of concept will provide the basis for the development of rapid test systems, e.g., for the detection of pathogens in water samples. KW - microfluidic paper analytic device (mu PAD) KW - patterning glass microfiber KW - fiber-electrophoresis chip KW - DNA KW - imprinted electrodes KW - cross layer chip KW - polymerase chain reaction (PCR) KW - purification Y1 - 2022 U6 - https://doi.org/10.3390/bios12020062 SN - 2079-6374 VL - 12 IS - 2 PB - MDPI CY - Basel ER - TY - THES A1 - Gonzalez Duran, Enrique T1 - Genetic control of intracellular gene transfer by DNA repair in N. tabacum N2 - Mitochondria and plastids are organelles with an endosymbiotic origin. During evolution, many genes are lost from the organellar genomes and get integrated in the nuclear genome, in what is known as intracellular/endosymbiotic gene transfer (IGT/EGT). IGT has been reproduced experimentally in Nicotiana tabacum at a gene transfer rate (GTR) of 1 event in 5 million cells, but, despite its centrality to eukaryotic evolution, there are no genetic factors known to influence the frequency of IGT in higher eukaryotes. The focus of this work was to determine the role of different DNA repair pathways of double strand break repair (DSBR) in the integration step of organellar DNA in the nuclear genome during IGT. Here, a CRISPR/Cas9 mutagenesis strategy was implemented in N. tabacum, with the aim of generating mutants in nuclear genes without expected visible phenotypes. This strategy led to the generation of a collection of independent mutants in the LIG4 (necessary for non-homologous end joining, NHEJ) and POLQ genes (necessary for microhomology mediated end joining, MMEJ). Targeting of other DSBR genes (KU70, KU80, RPA1C) generated mutants with unexpectedly strong developmental phenotypes.. These factors have telomeric roles, hinting towards a possible relationship between telomere length, and strength of developmental disruption upon loss of telomere structure in plants. The mutants were made in a genetic background encoding a plastid-encoded IGT reporter, that confers kanamycin resistance upon transfer to the nucleus. Through large scale independent experiments, increased IGT from the chloroplast to the nucleus was observed in lig4 mutants, as well as lines encoding a POLQ gene with a defective polymerase domain (polqΔPol). This shows that NHEJ or MMEJ have a double-sided relationship with IGT: while transferred genes may integrate using either pathway, the presence of both pathways suppresses IGT in wild-type somatic cells, thus demonstrating for the first time the extent on which nuclear genes control IGT frequency in plants. The IGT frequency increases in the mutants are likely mediated by increased availability of double strand breaks for integration. Additionally, kinetic analysis reveals that gene transfer (GT) events accumulate linearly as a function of time spent under antibiotic selection in the experiment, demonstrating that, contrary to what was previously thought, there is no such thing as a single GTR in somatic IGT experiments. Furthermore, IGT in tissue culture experiments appears to be the result of a "race against the clock" for integration in the nuclear genome, that starts when the organellar DNA arrives to the nucleus granting transient antibiotic resistance. GT events and escapes of kanamycin selection may be two possible outcomes from this race: those instances where the organellar DNA gets to integrate are recovered as GT events, and in those cases where timely integration fails, antibiotic resistance cannot be sustained, and end up considered as escapes. In the mutants, increased opportunities for integration in the nuclear genome change the overall ratio between IGT and escape events. The resources generated here are promising starting points for future research: (1) the mutant collection, for the further study of processes that depend on DNA repair in plants (2) the collection of GT lines obtained from these experiments, for the study of the effect of DSBR pathways over integration patterns and stability of transferred genes and (3) the developed CRISPR/Cas9 workflow for mutant generation, to make N. tabacum meet its potential as an attractive model for answering complex biological questions. N2 - Mitochondrien und Plastiden sind beides Organellen endosymbiotischen Ursprungs. Im Laufe der Evolution gehen viele Gene aus den Organellengenomen verloren und werden in das Kerngenom integriert, was als intrazellulärer/endosymbiotischer Gentransfer (IGT/EGT) bezeichnet wird. IGT konnte experimentell in Nicotiana tabacum mit einer Gentransferrate (GTR) von einem Ereignis in fünf Millionen Zellen nachgestellt werden, aber trotz seiner zentralen Bedeutung für die eukaryotische Evolution sind keine genetischen Faktoren bekannt, die die Häufigkeit von IGT in höheren Eukaryoten beeinflussen. Der Schwerpunkt dieser Arbeit lag auf der Bestimmung der Rolle verschiedener DNA-Reparaturwege der Doppelstrangbruchreparatur (DSBR) bei der Integration von Organellen-DNA in das Kerngenom während des IGT. Dazu wurde in N. tabacum eine CRISPR/Cas9-basierte Mutagenesestrategie angewandt, mit dem Ziel, Mutanten in Kerngenen zu erzeugen, für die keine sichtbaren Phänotypen zu erwarten sind. Diese Strategie führte zur Erzeugung einer Reihe unabhängiger Mutanten im LIG4-Gen (notwendig für „non-homologous end joining“, die nicht-homologe Verbindung von Enden, NHEJ) und POLQ (notwendig für „microhomology mediated end joining“, die Mikrohomologie-vermittelte Verbindung von Enden, MMEJ). Die gezielte Beeinflussung anderer DSBR-Gene (KU70, KU80, RPA1C) führte zu Mutanten mit unerwartet starken Entwicklungsphänotypen. Diese Gene spielen eine Rolle beim Erhalt der Telomere, was auf einen möglichen Zusammenhang zwischen der Telomerlänge und der Stärke der Entwicklungsstörung bei Verlust der Telomerstruktur in Pflanzen hindeutet. Die Mutanten wurden in einem genetischen Hintergrund erzeugt, der über einen in den Plastiden lokalisierten IGT-Reporter verfügt, der nach Übertragung in den Zellkern Kanamycin-Resistenz vermittelt. In groß angelegten unabhängigen Experimenten wurde in lig4-Mutanten sowie in Linien, die für ein POLQ-Gen mit einer defekten Polymerase-Domäne (polqΔPol) kodieren, eine erhöhte GTR vom Chloroplasten zum Zellkern beobachtet. Dies zeigt, dass NHEJ oder MMEJ eine zweischneidige Beziehung zum IGT haben: Während übertragene Gene folglich über jeden der beiden Mechanismen integriert werden können, unterdrückt das gleichzeitige Vorhandensein beider Wege IGT in somatischen Wildtyp-Zellen, wodurch zum ersten Mal gezeigt wird, in welchem Ausmaß Kerngene die IGT-Häufigkeit in Pflanzen kontrollieren. Die erhöhte Verfügbarkeit von Doppelstrangbrüchen für die Integration könnte für die erhöhte IGT-Häufigkeit in den Mutanten verantwortlich sein. Darüber hinaus zeigt die Analyse des Zeitverlaufs, dass Gentransferereignisse (GT) in Abhängigkeit von der Zeit, die im Experiment unter Antibiotikaselektion verbracht wurde, linear akkumulieren, was beweist, dass es, anders als bisher angenommen, in somatischen IGT-Experimenten keine statische GTR gibt. Darüber hinaus scheint IGT in Gewebekulturexperimenten das Ergebnis eines Wettlaufs mit der Zeit um die Integration in das Kerngenom zu sein, der beginnt, wenn die Organellen-DNA in den Zellkern gelangt und eine vorübergehende Antibiotikaresistenz gewährt. Echte GT-Ereignisse und „Escapes“ (scheinbare Resistenz, ein vorläufiges Ausweichen vor der Kanamycin-Selektion) können zwei mögliche Ergebnisse dieses Wettlaufs sein: Die Fälle, in denen die Organellen-DNA integriert wird, werden als GT-Ereignisse gewertet, und in den Fällen, in denen die rechtzeitige Integration scheitert, kann die Antibiotikaresistenz nicht aufrechterhalten werden und sie werden als „Escape“ betrachtet. In den Mutanten verändern sich die Möglichkeiten zur Integration in das Kerngenom, wodurch sich das Gesamtverhältnis zwischen IGT- und „Escape“-Ereignissen ändert. Die hier erzeugten Ressourcen sind vielversprechende Ausgangspunkte für künftige Forschungen: (1) die Mutantensammlung, für die Untersuchung von weiteren Prozessen, die von der DNA-Reparatur in Pflanzen abhängen, (2) die Sammlung von GT-Linien, die aus den hier beschriebenen Experimenten gewonnen wurden, für die Untersuchung der Auswirkungen von DSBR-Mechanismen auf Integrationsmuster und Stabilität übertragener Gene und (3) der hier entwickelte Arbeitsablauf für die Mutantenerzeugung mittels CRISPR/Cas9, damit N. tabacum sein Potenzial als attraktives Modell für die Beantwortung komplexer biologischer Fragestellungen erfüllen kann. T2 - Genetische Kontrolle des intrazellulären Gentransfers durch DNA-Reparatur in N. tabacum KW - endosymbiosis KW - organelles KW - gene KW - transfer KW - DNA KW - repair KW - genome KW - editing KW - evolution KW - plant Y1 - 2023 ER - TY - JOUR A1 - Fudickar, Werner A1 - Bauch, Marcel A1 - Ihmels, Heiko A1 - Linker, Torsten T1 - DNA-triggered enhancement of singlet oxygen production by pyridinium alkynylanthracenes JF - Chemistry - a European journal N2 - There is an ongoing interest in O-1(2) sensitizers, whose activity is selectively controlled by their interaction with DNA. To this end, we synthesized three isomeric pyridinium alkynylanthracenes 2 o-p and a water-soluble trapping reagent for O-1(2). In water and in the absence of DNA, these dyes show a poor efficiency to sensitize the photooxygenation of the trapping reagent as they decompose due to electron transfer processes. In contrast, in the presence of DNA O-1(2) is generated from the excited DNA-bound ligand. The interactions of 2 o-p with DNA were investigated by thermal DNA melting studies, UV/vis and fluorescence spectroscopy, and linear and circular dichroism spectroscopy. Our studies revealed an intercalative binding with an orientation of the long pyridyl-alkynyl axis parallel to the main axis of the DNA base pairs. In the presence of poly(dA : dT), all three isomers show an enhanced formation of singlet oxygen, as indicated by the reaction of the latter with the trapping reagent. With green light irradiation of isomer 2 o in poly(dA : dT), the conversion rate of the trapping reagent is enhanced by a factor >10. The formation of O-1(2) was confirmed by control experiments under anaerobic conditions, in deuterated solvents, or by addition of O-1(2) quenchers. When bound to poly(dG : dC), the opposite effect was observed only for isomers 2 o and 2 m, namely the trapping reagent reacted significantly slower. Overall, we showed that pyridinium alkynylanthracenes are very useful intercalators, that exhibit an enhanced photochemical O-1(2) generation in the DNA-bound state. KW - Anthracene KW - DNA KW - intercalations KW - photochemistry KW - singlet oxygen Y1 - 2021 U6 - https://doi.org/10.1002/chem.202101918 SN - 1521-3765 VL - 27 IS - 54 SP - 13591 EP - 13604 PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Cherstvy, Andrey G. A1 - Teif, Vladimir B. T1 - Electrostatic effect of H1-histone protein binding on nucleosome repeat length JF - Physical biology : a journal for the fundamental understanding of biological systems N2 - Within a simple biophysical model we describe the effect of electrostatic binding of H1 histone proteins on the nucleosome repeat length in chromatin. The length of wrapped DNA optimizes its binding energy to the histone core and the elastic energy penalty of DNA wrapping. The magnitude of the effect predicted from our model is in agreement with the systematic experimental data on the linear variation of nucleosome repeat lengths with H1/nucleosome ratio (Woodcock C L et al 2006 Chromos. Res. 14 17-25). We compare our model to the data for different cell types and organisms, with a widely varying ratio of bound H1 histones per nucleosome. We underline the importance of this non-specific histone-DNA charge-balance mechanism in regulating the positioning of nucleosomes and the degree of compaction of chromatin fibers in eukaryotic cells. KW - electrostatics KW - DNA KW - nucleosome Y1 - 2014 U6 - https://doi.org/10.1088/1478-3975/11/4/044001 SN - 1478-3967 SN - 1478-3975 VL - 11 IS - 4 PB - IOP Publ. Ltd. CY - Bristol ER - TY - GEN A1 - Beermann, Jan A1 - Westbury, Michael V. A1 - Hofreiter, Michael A1 - Hilgers, Leon A1 - Deister, Fabian A1 - Neumann, Hermann A1 - Raupach, Michael J. T1 - Cryptic species in a well-known habitat BT - applying taxonomics to the amphipod genus Epimeria (Crustacea, Peracarida) T2 - Postprints der Universität Potsdam : Mathematisch Naturwissenschaftliche Reihe N2 - Taxonomy plays a central role in biological sciences. It provides a communication system for scientists as it aims to enable correct identification of the studied organisms. As a consequence, species descriptions should seek to include as much available information as possible at species level to follow an integrative concept of 'taxonomics'. Here, we describe the cryptic species Epimeria frankei sp. nov. from the North Sea, and also redescribe its sister species, Epimeria cornigera. The morphological information obtained is substantiated by DNA barcodes and complete nuclear 18S rRNA gene sequences. In addition, we provide, for the first time, full mitochondrial genome data as part of a metazoan species description for a holotype, as well as the neotype. This study represents the first successful implementation of the recently proposed concept of taxonomics, using data from high-throughput technologies for integrative taxonomic studies, allowing the highest level of confidence for both biodiversity and ecological research. T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 1059 KW - multiple sequence alignment KW - Oxidase Subunit-I KW - mitochondrial genome KW - control region KW - Ribosomal-RNA KW - asellota crustacea KW - gammarus crustacea KW - deep-sea KW - DNA KW - evolution Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-460792 SN - 1866-8372 IS - 1059 ER - TY - JOUR A1 - Abouzar, Maryam H. A1 - Poghossian, Arshak A1 - Cherstvy, Andrey G. A1 - Pedraza, Angela M. A1 - Ingebrandt, Sven A1 - Schöning, Michael J. T1 - Label-free electrical detection of DNA by means of field-effect nanoplate capacitors experiments and modeling JF - Physica status solidi : A, Applications and materials science N2 - Label-free electrical detection of consecutive deoxyribonucleic acid (DNA) hybridization/denaturation by means of an array of individually addressable field-effect-based nanoplate silicon-on-insulator (SOI) capacitors modified with gold nanoparticles (Au-NP) is investigated. The proposed device detects charge changes on Au-NP/DNA hybrids induced by the hybridization or denaturation event. DNA hybridization was performed in a high ionic-strength solution to provide a high hybridization efficiency. On the other hand, to reduce the screening of the DNA charge by counter ions and to achieve a high sensitivity, the sensor signal induced by the hybridization and denaturation events was measured in a low ionic-strength solution. High sensor signals of about 120, 90, and 80 mV were registered after the DNA hybridization, denaturation, and re-hybridization events, respectively. Fluorescence microscopy has been applied as reference method to verify the DNA immobilization, hybridization, and denaturation processes. An electrostatic charge-plane model for potential changes at the gate surface of a nanoplate field-effect sensor induced by the DNA hybridization has been developed taking into account both the Debye length and the distance of the DNA charge from the gate surface. KW - DNA KW - field-effect KW - gold nanoparticle KW - label-free detection KW - nanoplate capacitor Y1 - 2012 U6 - https://doi.org/10.1002/pssa.201100710 SN - 1862-6300 VL - 209 IS - 5 SP - 925 EP - 934 PB - Wiley-VCH CY - Weinheim ER -