@phdthesis{Putzler2016, author = {Putzler, Sascha}, title = {Molekulare Charakterisierung des Centrosom-assoziierten Proteins CP91 in Dictyostelium discoideum}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-394689}, school = {Universit{\"a}t Potsdam}, pages = {111}, year = {2016}, abstract = {Das Dictyostelium-Centrosom ist ein Modell f{\"u}r acentriol{\"a}re Centrosomen. Es besteht aus einer dreischichtigen Kernstruktur und ist von einer Corona umgeben, welche Nukleationskomplexe f{\"u}r Mikrotubuli beinhaltet. Die Verdoppelung der Kernstruktur wird einmal pro Zellzyklus am {\"U}bergang der G2 zur M-Phase gestartet. Durch eine Proteomanalyse isolierter Centrosomen konnte CP91 identifiziert werden, ein 91 kDa großes Coiled-Coil Protein, das in der centrosomalen Kernstruktur lokalisiert. GFP-CP91 zeigte fast keine Mobilit{\"a}t in FRAP-Experimenten w{\"a}hrend der Interphase, was darauf hindeutet, dass es sich bei CP91 um eine Strukturkomponente des Centrosoms handelt. In der Mitose hingegen dissoziieren das GFP-CP91 als auch das endogene CP91 ab und fehlen an den Spindelpolen von der sp{\"a}ten Prophase bis zur Anaphase. Dieses Verhalten korreliert mit dem Verschwinden der zentralen Schicht der Kernstruktur zu Beginn der Centrosomenverdopplung. Somit ist CP91 mit großer Wahrscheinlichkeit ein Bestandteil dieser Schicht. CP91-Fragmente der N-terminalen bzw. C-terminalen Dom{\"a}ne (GFP-CP91 N-Terminus, GFP-CP91 C-Terminus) lokalisieren als GFP-Fusionsproteine exprimiert auch am Centrosom, zeigen aber nicht die gleiche mitotische Verteilung des Volll{\"a}ngenproteins. Das CP91-Fragment der zentralen Coiled-Coil Dom{\"a}ne (GFP-CP91cc) lokalisiert als GFP-Fusionsprotein exprimiert, als ein diffuser cytosolische Cluster, in der N{\"a}he des Centrosoms. Es zeigt eine partiell {\"a}hnliche mitotische Verteilung wie das Volll{\"a}ngenprotein. Dies l{\"a}sst eine regulatorische Dom{\"a}ne innerhalb der Coiled-Coil Dom{\"a}ne vermuten. Die Expression der GFP-Fusionsproteine unterdr{\"u}ckt die Expression des endogenen CP91 und bringt {\"u}berz{\"a}hlige Centrosomen hervor. Dies war auch eine markante Eigenschaft nach der Unterexpression von CP91 durch RNAi. Zus{\"a}tzlich zeigte sich in CP91-RNAi Zellen eine stark erh{\"o}hte Ploidie verursacht durch schwere Defekte in der Chromosomensegregation verbunden mit einer erh{\"o}hten Zellgr{\"o}ße und Defekten im Abschn{\"u}rungsprozess w{\"a}hrend der Cytokinese. Die Unterexpression von CP91 durch RNAi hatte auch einen direkten Einfluss auf die Menge an den centrosomalen Proteinen CP39, CP55 und CEP192 und dem Centromerprotein Cenp68 in der Interphase. Die Ergebnisse deuten darauf hin, dass CP91 eine zentrale centrosomale Kernkomponente ist und f{\"u}r den Zusammenhalt der beiden {\"a}ußeren Schichten der Kernstruktur ben{\"o}tigt wird. Zudem spielt CP91 eine wichtige Rolle f{\"u}r eine ordnungsgem{\"a}ße Centrosomenbiogenese und, unabh{\"a}ngig davon, bei dem Abschn{\"u}rungsprozess der Tochterzellen w{\"a}hrend der Cytokinese.}, language = {de} } @phdthesis{BeineGolovchuk2016, author = {Beine-Golovchuk, Olga}, title = {Characterization and functional complementation of the arabidopsis ribosomal Reil1 - 1Reil2-1 double mutant}, school = {Universit{\"a}t Potsdam}, pages = {106}, year = {2016}, language = {en} } @phdthesis{RuizMartinez2016, author = {Ruiz-Martinez, Maria}, title = {Characterisation and engineering of lignocellulolytic enzymes from the soil bacterium Sorangium cellulosum}, school = {Universit{\"a}t Potsdam}, pages = {137}, year = {2016}, language = {en} } @phdthesis{Geyer2016, author = {Geyer, Juliane}, title = {Adapting biodiversity conservation management to climate change}, school = {Universit{\"a}t Potsdam}, pages = {198}, year = {2016}, language = {en} } @phdthesis{Connor2016, author = {Connor, Daniel Oliver}, title = {Identifikation und Charakterisierung neuer immunogener Proteine und anschließende Generierung rekombinanter Antik{\"o}rper mittels Phage Display}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-104120}, school = {Universit{\"a}t Potsdam}, pages = {VII, 112, lv Seiten}, year = {2016}, abstract = {Seit der Einf{\"u}hrung von Antibiotika in die medizinische Behandlung von bakteriellen Infektionskrankheiten existiert ein Wettlauf zwischen der Evolution von Bakterienresistenzen und der Entwicklung wirksamer Antibiotika. W{\"a}hrend bis in die 80er Jahre verst{\"a}rkt an neuen Antibiotika geforscht wurde, gewinnen multiresistente Keime heute zunehmend die Oberhand. Um einzelne Pathogene erfolgreich nachzuweisen und zu bek{\"a}mpfen, ist ein grundlegendes Wissen {\"u}ber den Erreger unumg{\"a}nglich. Bakterielle Proteine, die bei einer Infektion vorrangig vom Immunsystem prozessiert und pr{\"a}sentiert werden, k{\"o}nnten f{\"u}r die Entwicklung von Impfstoffen oder gezielten Therapeutika n{\"u}tzlich sein. Auch f{\"u}r die Diagnostik w{\"a}ren diese immundominanten Proteine interessant. Allerdings herrscht ein Mangel an Wissen {\"u}ber spezifische Antigene vieler pathogener Bakterien, die eine eindeutige Diagnostik eines einzelnen Erregers erlauben w{\"u}rden. Daher wurden in dieser Arbeit vier verschiedene Humanpathogene mittels Phage Display untersucht: Neisseria gonorrhoeae, Neisseria meningitidis, Borrelia burgdorferi und Clostridium difficile. Hierf{\"u}r wurden aus der genomischen DNA der vier Erreger Bibliotheken konstruiert und durch wiederholte Selektion und Amplifikation, dem sogenannten Panning, immunogene Proteine isoliert. F{\"u}r alle Erreger bis auf C. difficile wurden immunogene Proteine aus den jeweiligen Bibliotheken isoliert. Die identifizierten Proteine von N. meningitidis und B. burgdorferi waren gr{\"o}ßtenteils bekannt, konnten aber in dieser Arbeit durch Phage Display verifiziert werden. F{\"u}r N. gonorrhoeae wurden 21 potentiell immunogene Oligopeptide isoliert, von denen sechs Proteine als neue zuvor unbeschriebene Proteine mit immunogenem Charakter identifiziert wurden. Von den Phagen-pr{\"a}sentierten Oligopeptide der 21 immunogenen Proteine wurden Epitopmappings mit verschiedenen polyklonalen Antik{\"o}rpern durchgef{\"u}hrt, um immunogene Bereiche n{\"a}her zu identifizieren und zu charakterisieren. Bei zehn Proteinen wurden lineare Epitope eindeutig mit drei polyklonalen Antik{\"o}rpern identifiziert, von f{\"u}nf weiteren Proteinen waren Epitope mit mindestens einem Antik{\"o}rper detektierbar. F{\"u}r eine weitere Charakterisierung der ermittelten Epitope wurden Alaninscans durchgef{\"u}hrt, die eine detaillierte Auskunft {\"u}ber kritische Aminos{\"a}uren f{\"u}r die Bindung des Antik{\"o}rpers an das Epitop geben. Ausgehend von dem neu identifizierten Protein mit immunogenem Charakter NGO1634 wurden 26 weitere Proteine aufgrund ihrer funktionellen {\"A}hnlichkeit ausgew{\"a}hlt und mithilfe bioinformatischer Analysen auf ihre Eignung zur Entwicklung einer diagnostischen Anwendung analysiert. Durch Ausschluss der meisten Proteine aufgrund ihrer Lokalisation, Membrantopologie oder unspezifischen Proteinsequenz wurden scFv-Antik{\"o}rper gegen acht Proteine mittels Phage Display generiert und anschließend als scFv-Fc-Fusionsantik{\"o}rper produziert und charakterisiert. Die hier identifizierten Proteine und linearen Epitope k{\"o}nnten einen Ansatzpunkt f{\"u}r die Entwicklung einer diagnostischen oder therapeutischen Anwendung bieten. Lineare Epitopsequenzen werden h{\"a}ufig f{\"u}r die Impfstoffentwicklung eingesetzt, sodass vor allem die in dieser Arbeit bestimmten Epitope von Membranproteinen interessante Kandidaten f{\"u}r weitere Untersuchungen in diese Richtung sind. Durch weitere Untersuchungen k{\"o}nnten m{\"o}glicherweise unbekannte Virulenzfaktoren entdeckt werden, deren Inhibierung einen entscheidenden Einfluss auf Infektionen haben k{\"o}nnten.}, language = {de} } @phdthesis{Heinze2016, author = {Heinze, Johannes}, title = {The impact of soil microbiota on plant species performance and diversity in semi-natural grasslands}, school = {Universit{\"a}t Potsdam}, pages = {157}, year = {2016}, language = {en} } @phdthesis{Reichel2016, author = {Reichel, Victoria Eleonore}, title = {Biomedical applications and multifunctional nanostructures based on magnetite nanoparticles synthesized in presence of biological additives}, school = {Universit{\"a}t Potsdam}, pages = {94, XX}, year = {2016}, language = {en} } @phdthesis{OnanaEloundouEpseMbebi2016, author = {Onana Eloundou Epse Mbebi, Jeanne Marie}, title = {Robustness and plasticity in chemical reaction networks}, school = {Universit{\"a}t Potsdam}, pages = {116}, year = {2016}, language = {en} } @phdthesis{Fiedler2016, author = {Fiedler, Dorothea}, title = {Impact of Dissolved Organic Nitrogen on Freshwater Phytoplankton}, school = {Universit{\"a}t Potsdam}, pages = {XIII, 92}, year = {2016}, abstract = {In freshwater sciences, nitrogen gained increasing attention in the past as an important resource potentially influencing phytoplankton growth and thus eutrophication. Most studies and all management approaches, however, are still restricted to dissolved inorganic nitrogen (DIN = nitrate + nitrite + ammonium) since dissolved organic nitrogen (DON) was considered to be refractory for most of the photoautotrophs. In the meantime this assumption has been disproved for all aquatic systems. While research on DON in marine ecosystems substantially increased, in freshwater a surprisingly small number of investigations has been carried out on DON utilization by phytoplankton or even the occurrence and seasonal development of total DON or its compounds in lakes. Therefore, our present knowledge on DON utilization by phytoplankton is often based on single species experiments using a sole, usually low molecular weight DON component, often in unnaturally high amounts mainly carried out with marine phytoplankton species. Thus, we know that some phytoplankton species can take up different DON fractions if they are available in high concentrations and as sole nitrogen source. This does not necessarily imply that phytoplankton would perform likewise in natural environments. In addition, it will be difficult to draw conclusions on the behavior of freshwater phytoplankton from experiments with marine phytoplankton since the nutrient regime in marine environments differs from that of freshwater. In the light of the parallel availability of inorganic and organic nitrogen species in natural freshwater ecosystems, several questions must be raised: "If inorganic nitrogen is available, would phytoplankton really rely on an organic nitrogen source? Could a connection be detected between the seasonal development of DON and changes in the phytoplankton community composition as found for inorganic nitrogen? And if we reduce the input of inorganic nitrogen in lakes and rivers would the importance of DON as nitrogen source for phytoplankton increase, counteracting all management efforts or even leading to undesired effects due to changes in phytoplankton physiology and biodiversity?" I experimentally addressed the questions whether those DON compounds differentially influence growth, physiology and composition of phytoplankton both as sole available nitrogen source and in combination with other nitrogen compounds. I hypothesized that all offered DON - compounds (urea, natural organic matter (NOM), dissolved free and combined amino acids (DFAA, DCAA)) could be utilized by phytoplankton at natural concentrations. However, I assumed that the availability would decrease with increasing compound complexity. I furthermore hypothesized that the occurrence of low DIN concentrations would not affect the utilization of DON negatively. The nitrogen source, whatsoever, would have an impact on phytoplankton physiology as well as community composition. To investigate these questions and assumptions I conducted bioassays with algae monocultures as well as phytoplankton communities testing the utilization of various DON compounds by several freshwater phytoplankton species. Especially the potential utilization of NOM, a complex DON compound mainly consisting of humic substances is of interest, since it is usually regarded to be refractory. In order to be able to use natural concentrations of DON - compounds for my experiments the concentration of total DON and some DON - compounds (urea, humic substances, heigh molecular weight substances) was assessed in Lake M{\"u}ggelsee. All compounds were able to support algae growth in the low natural concentrations supplied. However, I found that the offered DON compounds differ in their availability to various algae species, both, as sole nitrogen source or in combination with low DIN concentrations. As expected, the availability decreased with increasing complexity of the nitrogen compound. Furthermore, I could show that changes in algal physiology (nitrogen storage, metabolism) occur depending on the utilized nitrogen source. Especially the secondary photosynthetic pigment composition, heterocyst frequency and C:N - ratio of the algae were affected. The uptake and usage of certain nitrogen compounds might be more costly, potentially resulting in those physiology changes. Whereas laboratory experiments with single species revealed strong effects of DON, algal responses to DON in a multi-species situation remain unclear. Experiments with phytoplankton communities from Lake M{\"u}ggelsee revealed that the nitrogen pool composition does influence the phytoplankton community structure. The findings furthermore show that several species combined might utilize the supplied nitrogen completely different than monocultures in the laboratory. Thus, besides the actual ability of algae to use the offered nitrogen sources other factors, such as interspecific competition, may be of importance. I further investigated, if the results of the laboratory experiments, can be verified in the field. Here, I surveyed the seasonal development of several dissolved organic matter (DOM) components (urea, high molecular weight substances (HMWS), humic substances (HS)) and associated parameters (Specific UV-absorption (SUVA), C:N - ratio) in Lake M{\"u}ggelsee between 2011 and 2013. Furthermore, data from the long term measurements series of Lake M{\"u}ggelsee such as physical (temperature, light, pH, O2) and chemical parameters (nitrogen, phosphorous, silica, inorganic carbon), zooplankton and phytoplankton data were used to investigate how much of the variability of the phytoplankton composition in Lake M{\"u}ggelsee can be explained by DON/DOM concentration and composition, relative to the other groups of explanatory variables. The results show that DON mainly consists of rather complex compounds such as humic substances and biopolymers (80 \%) and that only slight seasonal trends are detectable. Using variance partitioning I could show, that the usually investigated nutrients (DIN, silica, inorganic carbon, phosphorous) and abiotic factors together explain most of the algae composition as was to be expected (57.1 \% of modeled variance). However, DOM and the associated parameters uniquely explain 10.3 \% of the variance and thus slightly more than zooplankton with 9.3 \%. I could therefore prove, that the composition of DOM (nitrogen and carbon) is connected to the algae composition in an eutrophic lake such as Lake M{\"u}ggelsee. DON - compounds such as urea, however, could not be correlated with the occurrence of specific phytoplankton species. Overall, the results of this study imply that DON can be a valuable nitrogen source for freshwater phytoplankton. DON is used by various species even when DIN is available in low concentrations. Through the reduction of DIN in lakes and rivers, the DON:DIN ratio might be changed, resulting even in an increased importance of DON as phytoplankton nitrogen source. My work suggests that not only N2-fixation but also DON utilization might compensate for reduced N - input. Changes from DIN to DON as main nitrogen source might also promote certain, potentially undesired algae species and influence the biodiversity of a limnic ecosystem through changes in the phytoplankton community structure. Thus, DON, especially urea, should be included in calculations concerning total available nitrogen and when determining nitrogen threshold values. Furthermore, the input-reduction of DON, for example from waste-water treatment plants should also be evaluated and the results of my thesis should find consideration when planning to reduce the nitrogen input in freshwater.}, language = {en} } @phdthesis{EbrahimianMotlagh2016, author = {Ebrahimian Motlagh, Saghar}, title = {Functional characterization of stress-responsive transcription factors and their gene regulatory networks in Arabidopsis thaliana}, school = {Universit{\"a}t Potsdam}, pages = {155, X}, year = {2016}, language = {en} }