@misc{JonesGonzalezFortesConnelletal.2015, author = {Jones, Eppie R. and Gonz{\´a}lez-Fortes, Gloria M. and Connell, Sarah and Siska, Veronika and Eriksson, Anders and Martiniano, Rui and McLaughlin, Russell L. and Llorente, Marcos Gallego and Cassidy, Lara M. and Gamba, Cristina and Meshveliani, Tengiz and Bar-Yosef, Ofer and M{\"u}ller, Werner and Belfer-Cohen, Anna and Matskevich, Zinovi and Jakeli, Nino and Higham, Thomas F. G. and Currat, Mathias and Lordkipanidze, David and Hofreiter, Michael and Manica, Andrea and Pinhasi, Ron and Bradley, Daniel G.}, title = {Upper Palaeolithic genomes reveal deep roots of modern Eurasians}, series = {Zweitver{\"o}ffentlichungen der Universit{\"a}t Potsdam : Mathematisch-Naturwissenschaftliche Reihe}, journal = {Zweitver{\"o}ffentlichungen der Universit{\"a}t Potsdam : Mathematisch-Naturwissenschaftliche Reihe}, number = {1334}, issn = {1866-8372}, doi = {10.25932/publishup-43931}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-439317}, pages = {8}, year = {2015}, abstract = {We extend the scope of European palaeogenomics by sequencing the genomes of Late Upper Palaeolithic (13,300 years old, 1.4-fold coverage) and Mesolithic (9,700 years old, 15.4-fold) males from western Georgia in the Caucasus and a Late Upper Palaeolithic (13,700 years old, 9.5-fold) male from Switzerland. While we detect Late Palaeolithic-Mesolithic genomic continuity in both regions, we find that Caucasus hunter-gatherers (CHG) belong to a distinct ancient clade that split from western hunter-gatherers ∼45 kya, shortly after the expansion of anatomically modern humans into Europe and from the ancestors of Neolithic farmers ∼25 kya, around the Last Glacial Maximum. CHG genomes significantly contributed to the Yamnaya steppe herders who migrated into Europe ∼3,000 BC, supporting a formative Caucasus influence on this important Early Bronze age culture. CHG left their imprint on modern populations from the Caucasus and also central and south Asia possibly marking the arrival of Indo-Aryan languages.}, language = {en} } @misc{HartmannHasenkampMayeretal.2015, author = {Hartmann, Stefanie and Hasenkamp, Natascha and Mayer, Jens and Michaux, Johan and Morand, Serge and Mazzoni, Camila J. and Roca, Alfred L. and Greenwood, Alex D.}, title = {Endogenous murine leukemia retroviral variation across wild European and inbred strains of house mouse}, series = {Zweitver{\"o}ffentlichungen der Universit{\"a}t Potsdam : Mathematisch-Naturwissenschaftliche Reihe}, journal = {Zweitver{\"o}ffentlichungen der Universit{\"a}t Potsdam : Mathematisch-Naturwissenschaftliche Reihe}, number = {1329}, issn = {1866-8372}, doi = {10.25932/publishup-43120}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-431200}, pages = {13}, year = {2015}, abstract = {Background: Endogenous murine leukemia retroviruses (MLVs) are high copy number proviral elements difficult to comprehensively characterize using standard low throughput sequencing approaches. However, high throughput approaches generate data that is challenging to process, interpret and present. Results: Next generation sequencing (NGS) data was generated for MLVs from two wild caught Mus musculus domesticus (from mainland France and Corsica) and for inbred laboratory mouse strains C3H, LP/J and SJL. Sequence reads were grouped using a novel sequence clustering approach as applied to retroviral sequences. A Markov cluster algorithm was employed, and the sequence reads were queried for matches to specific xenotropic (Xmv), polytropic (Pmv) and modified polytropic (Mpmv) viral reference sequences. Conclusions: Various MLV subtypes were more widespread than expected among the mice, which may be due to the higher coverage of NGS, or to the presence of similar sequence across many different proviral loci. The results did not correlate with variation in the major MLV receptor Xpr1, which can restrict exogenous MLVs, suggesting that endogenous MLV distribution may reflect gene flow more than past resistance to infection.}, language = {en} } @misc{ElsnerSchiblerHofreiteretal.2015, author = {Elsner, Julia and Schibler, J{\"o}rg and Hofreiter, Michael and Schlumbaum, Angela}, title = {Burial condition is the most important factor for mtDNA PCR amplification success in Palaeolithic equid remains from the Alpine foreland}, series = {Postprints der Universit{\"a}t Potsdam Mathematisch-Naturwissenschaftliche Reihe}, journal = {Postprints der Universit{\"a}t Potsdam Mathematisch-Naturwissenschaftliche Reihe}, number = {727}, issn = {1866-8372}, doi = {10.25932/publishup-42976}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-429763}, pages = {505 -- 515}, year = {2015}, abstract = {Faunal remains from Palaeolithic sites are important genetic sources to study preglacial and postglacial populations and to investigate the effect of climate change and human impact. Post mortem decay, resulting in fragmented and chemically modified DNA, is a key obstacle in ancient DNA analyses. In the absence of reliable methods to determine the presence of endogenous DNA in sub-fossil samples, temporal and spatial surveys of DNA survival on a regional scale may help to estimate the potential of faunal remains from a given time period and region. We therefore investigated PCR amplification success, PCR performance and post mortem damage in c. 47,000 to c. 12,000-year-old horse remains from 14 Palaeolithic sites along the Swiss Jura Mountains in relation to depositional context, tissue type, storage time and age, potentially influencing DNA preservation. The targeted 75 base pair mitochondrial DNA fragment could be amplified solely from equid remains from caves and not from any of the open dry and (temporary) wetland sites. Whether teeth are better than bones cannot be ultimately decided; however, both storage time after excavation and age significantly affect PCR amplification and performance, albeit not in a linear way. This is best explained by the—inevitable—heterogeneity of the data set. The extent of post mortem damage is not related to any of the potential impact factors. The results encourage comprehensive investigations of Palaeolithic cave sites, even from temperate regions.}, language = {en} } @misc{UestuenBartetzkoBoernke2015, author = {{\"U}st{\"u}n, Suayib and Bartetzko, Verena and B{\"o}rnke, Frederik}, title = {The Xanthomonas effector XopJ triggers a conditional hypersensitive response upon treatment of N. benthamiana leaves with salicylic acid}, series = {Frontiers in plant science}, journal = {Frontiers in plant science}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-406537}, pages = {11}, year = {2015}, abstract = {XopJ is a Xanthomonas type III effector protein that promotes bacterial virulence on susceptible pepper plants through the inhibition of the host cell proteasome and a resultant suppression of salicylic acid (SA) - dependent defense responses. We show here that Nicotiana benthamiana leaves transiently expressing XopJ display hypersensitive response (HR) -like symptoms when exogenously treated with SA. This apparent avirulence function of XopJ was further dependent on effector myristoylation as well as on an intact catalytic triad, suggesting a requirement of its enzymatic activity for HR-like symptom elicitation. The ability of XopJ to cause a HR-like symptom development upon SA treatment was lost upon silencing of SGT1 and NDR1, respectively, but was independent of EDS1 silencing, suggesting that XopJ is recognized by an R protein of the CC-NBS-LRR class. Furthermore, silencing of NPR1 abolished the elicitation of HR-like symptoms in XopJ expressing leaves after SA application. Measurement of the proteasome activity indicated that proteasome inhibition by XopJ was alleviated in the presence of SA, an effect that was not observed in NPR1 silenced plants. Our results suggest that XopJ - triggered HR-like symptoms are closely related to the virulence function of the effector and that XopJ follows a two-signal model in order to elicit a response in the non-host plant N. benthamiana.}, language = {en} } @misc{KehrDittmannThuenemann2015, author = {Kehr, Jan-Christoph and Dittmann-Th{\"u}nemann, Elke}, title = {Biosynthesis and function of extracellular glycans in cyanobacteria}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-400121}, pages = {17}, year = {2015}, abstract = {The cell surface of cyanobacteria is covered with glycans that confer versatility and adaptability to a multitude of environmental factors. The complex carbohydrates act as barriers against different types of stress and play a role in intra- as well as inter-species interactions. In this review, we summarize the current knowledge of the chemical composition, biosynthesis and biological function of exo- and lipo-polysaccharides from cyanobacteria and give an overview of sugar-binding lectins characterized from cyanobacteria. We discuss similarities with well-studied enterobacterial systems and highlight the unique features of cyanobacteria. We pay special attention to colony formation and EPS biosynthesis in the bloom-forming cyanobacterium, Microcystis aeruginosa.}, language = {en} } @misc{McKennaLeimkuehlerHerteretal.2015, author = {McKenna, Shane M. and Leimk{\"u}hler, Silke and Herter, Susanne and Turner, Nicholas J. and Carnell, Andrew J.}, title = {Enzyme cascade reactions}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-102271}, pages = {3271 -- 3275}, year = {2015}, abstract = {A one-pot tandem enzyme reaction using galactose oxidase M3-5 and aldehyde oxidase PaoABC was used to convert hydroxymethylfurfural (HMF) to the pure bioplastics precursor FDCA in 74\% isolated yield. A range of alcohols was also converted to carboxylic acids in high yield under mild conditions.}, language = {en} } @misc{MuinodeBruijnPajoroetal.2015, author = {Mui{\~n}o, Jose M. and de Bruijn, Suzanne and Pajoro, Alice and Geuten, Koen and Vingron, Martin and Angenent, Gerco C. and Kaufmann, Kerstin}, title = {Evolution of DNA-Binding Sites of a Floral Master Regulatory Transcription Factor}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-96580}, pages = {1225 -- 1245}, year = {2015}, abstract = {Flower development is controlled by the action of key regulatory transcription factors of the MADS-domain family. The function of these factors appears to be highly conserved among species based on mutant phenotypes. However, the conservation of their downstream processes is much less well understood, mostly because the evolutionary turnover and variation of their DNA-binding sites (BSs) among plant species have not yet been experimentally determined. Here, we performed comparative ChIP (chromatin immunoprecipitation)-seq experiments of the MADS-domain transcription factor SEPALLATA3 (SEP3) in two closely related Arabidopsis species: Arabidopsis thaliana and A. lyrata which have very similar floral organ morphology. We found that BS conservation is associated with DNA sequence conservation, the presence of the CArG-box BS motif and on the relative position of the BS to its potential target gene. Differences in genome size and structure can explain that SEP3 BSs in A. lyrata can be located more distantly to their potential target genes than their counterparts in A. thaliana. In A. lyrata, we identified transposition as a mechanism to generate novel SEP3 binding locations in the genome. Comparative gene expression analysis shows that the loss/gain of BSs is associated with a change in gene expression. In summary, this study investigates the evolutionary dynamics of DNA BSs of a floral key-regulatory transcription factor and explores factors affecting this phenomenon.}, language = {en} } @misc{KappelTrostCzesnicketal.2015, author = {Kappel, Christian and Trost, Gerda and Czesnick, Hj{\"o}rdis and Ramming, Anna and Kolbe, Benjamin and Vi, Song Lang and Bispo, Cl{\´a}udia and Becker, J{\"o}rg D. and de Moor, Cornelia and Lenhard, Michael}, title = {Genome-Wide Analysis of PAPS1-Dependent Polyadenylation Identifies Novel Roles for Functionally Specialized Poly(A) Polymerases in Arabidopsis thaliana}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-96400}, pages = {1 -- 30}, year = {2015}, abstract = {The poly(A) tail at 3' ends of eukaryotic mRNAs promotes their nuclear export, stability and translational efficiency, and changes in its length can strongly impact gene expression. The Arabidopsis thaliana genome encodes three canonical nuclear poly(A) polymerases, PAPS1, PAPS2 and PAPS4. As shown by their different mutant phenotypes, these three isoforms are functionally specialized, with PAPS1 modifying organ growth and suppressing a constitutive immune response. However, the molecular basis of this specialization is largely unknown. Here, we have estimated poly(A)-tail lengths on a transcriptome-wide scale in wild-type and paps1 mutants. This identified categories of genes as particularly strongly affected in paps1 mutants, including genes encoding ribosomal proteins, cell-division factors and major carbohydrate-metabolic proteins. We experimentally verified two novel functions of PAPS1 in ribosome biogenesis and redox homoeostasis that were predicted based on the analysis of poly(A)-tail length changes in paps1 mutants. When overlaying the PAPS1-dependent effects observed here with coexpression analysis based on independent microarray data, the two clusters of transcripts that are most closely coexpressed with PAPS1 show the strongest change in poly(A)-tail length and transcript abundance in paps1 mutants in our analysis. This suggests that their coexpression reflects at least partly the preferential polyadenylation of these transcripts by PAPS1 versus the other two poly(A)-polymerase isoforms. Thus, transcriptome-wide analysis of poly(A)-tail lengths identifies novel biological functions and likely target transcripts for polyadenylation by PAPS1. Data integration with large-scale co-expression data suggests that changes in the relative activities of the isoforms are used as an endogenous mechanism to co-ordinately modulate plant gene expression.}, language = {en} } @masterthesis{Kreibich2015, type = {Bachelor Thesis}, author = {Kreibich, Christoph}, title = {Erucas{\"a}ure in Brassica napus L. - ein ph{\"a}notypisches Merkmal im Genetikunterricht und ihr Nachweis mit Hilfe von Papierchromatographie}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-93341}, school = {Universit{\"a}t Potsdam}, pages = {II, 35}, year = {2015}, abstract = {Erucic acid is a mono-unsaturated fatty acid that is naturally found in large quantities in seeds of rapeseed (Brassica napus L.) and other Brassica species. Erucic acid represents an important resource in the industry, however, due to its injurious effects on the heart muscle, this fatty acid is considered to be nutritionally harmful. Therefore, new high quality rapeseed cultivars were bred in order to eliminate the content of erucic acid in rapeseed oil at the end of the 20th century. In the breeding process, paper chromatography was used for the distinction between seeds with high and low content of erucic acid. Here, this outdated method was revised and optimized for educational purposes. By means of paper chromatography the qualitative content of erucic acid and four other unsaturated fatty acids was analyzed in rapeseed and linseed. The character 'erucic acid content', determined by two additive genes, can be used as a practical example of a phenotypic marker in school lessons, for instance, in the course 'achievement of plant breeding'. Thus, this qualitative analysis of erucic acid content enables the teacher to connect classical genetics with modern methods of plant genetics.}, language = {de} } @misc{SicardKappelJosephsetal.2015, author = {Sicard, Adrien and Kappel, Christian and Josephs, Emily B. and Wha Lee, Young and Marona, Cindy and Stinchcombe, John R. and Wright, Stephen I. and Lenhard, Michael}, title = {Divergent sorting of a balanced ancestral polymorphism underlies the establishment of gene-flow barriers in Capsella}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-93568}, year = {2015}, abstract = {In the Bateson-Dobzhansky-Muller model of genetic incompatibilities post-zygotic gene-flow barriers arise by fixation of novel alleles at interacting loci in separated populations. Many such incompatibilities are polymorphic in plants, implying an important role for genetic drift or balancing selection in their origin and evolution. Here we show that NPR1 and RPP5 loci cause a genetic incompatibility between the incipient species Capsella grandiflora and C. rubella, and the more distantly related C. rubella and C. orientalis. The incompatible RPP5 allele results from a mutation in C. rubella, while the incompatible NPR1 allele is frequent in the ancestral C. grandiflora. Compatible and incompatible NPR1 haplotypes are maintained by balancing selection in C. grandiflora, and were divergently sorted into the derived C. rubella and C. orientalis. Thus, by maintaining differentiated alleles at high frequencies, balancing selection on ancestral polymorphisms can facilitate establishing gene-flow barriers between derived populations through lineage sorting of the alternative alleles.}, language = {en} } @misc{ValentePhillimoreEtienne2015, author = {Valente, Luis M. and Phillimore, Albert B. and Etienne, Rampal S.}, title = {Equilibrium and non-equilibrium dynamics simultaneously operate in the Gal{\´a}pagos islands}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-93525}, pages = {9}, year = {2015}, abstract = {Island biotas emerge from the interplay between colonisation, speciation and extinction and are often the scene of spectacular adaptive radiations. A common assumption is that insular diversity is at a dynamic equilibrium, but for remote islands, such as Hawaii or Gal{\´a}pagos, this idea remains untested. Here, we reconstruct the temporal accumulation of terrestrial bird species of the Gal{\´a}pagos using a novel phylogenetic method that estimates rates of biota assembly for an entire community. We show that species richness on the archipelago is in an ascending phase and does not tend towards equilibrium. The majority of the avifauna diversifies at a slow rate, without detectable ecological limits. However, Darwin's finches form an exception: they rapidly reach a carrying capacity and subsequently follow a coalescent-like diversification process. Together, these results suggest that avian diversity of remote islands is rising, and challenge the mutual exclusivity of the non-equilibrium and equilibrium ecological paradigms.}, language = {en} } @phdthesis{Liebrich2015, author = {Liebrich, Marietta}, title = {Einfluss von Prozessoptimierungen auf die mikrobielle Diversit{\"a}t und die Effizienz der Gasbildung in Co-Verg{\"a}rungsanlagen der Abfallwirtschaft}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-91066}, school = {Universit{\"a}t Potsdam}, pages = {VII, 102}, year = {2015}, abstract = {Im Hinblick auf die Problematik der Umweltverschmutzung durch die Nutzung fossiler Brennstoffe ist es n{\"o}tig, eine langfristig stabile und umweltfreundliche Energieversorgung zu gew{\"a}hrleisten. Eine M{\"o}glichkeit, den Energiebedarf CO2-neutral zu decken, ist die Nutzung von Biogas. Hierbei spielt der Einsatz von biogenen Reststoffen, die durch einen hohen Anteil an Kohlenhydraten, Fetten und Proteinen gekennzeichnet sind und daher ein hohes Biogaspotential besitzen, eine wichtige Rolle. Voraussetzung f{\"u}r die Effizienz und Rentabilit{\"a}t solcher Anlagen ist u. a. ein stabiler Gasbildungsprozess. Da bisher noch nicht alle Aspekte der Biogasbildung vollst{\"a}ndig verstanden sind, werden die Anlagen oft nicht optimal ausgelastet, um Prozessst{\"o}rungen wie z. B. {\"U}bers{\"a}uerung zu vermeiden. Um dennoch auftretende Prozessst{\"o}rungen zu beheben, k{\"o}nnen unterschiedliche Maßnahmen durchgef{\"u}hrt werden. Neben der Senkung der Raumbelastung, ist es m{\"o}glich, den pH-Wert durch die Zugabe von Natronlauge oder Calciumoxid anzuheben. In der vorliegenden Arbeit wurden sowohl Prozessst{\"o}rungen als auch Prozessregenerierungen an einer großtechnischen Biogasanlage und in Laborversuchen untersucht. Dabei galt es, neben den physikalischen und chemischen Parametern, die mikrobielle Bioz{\"o}nose mit Hilfe des genetischen Fingerprintings zu charakterisieren und {\"A}nderungen zu detektieren. W{\"a}hrend der Prozessregenerierungen wurden nach der Zugabe von CaO Ver{\"a}nderungen des G{\"a}rrestes beobachtet. Es bildeten sich Pellets, die im Hinblick auf ihre Funktion f{\"u}r die Prozessregenerierung und die Prozessstabilit{\"a}t molekularbiologisch und mikroskopisch untersucht wurden. Es wurde weiterhin der Frage nachgegangen, welche Rolle die Mikroorganismen bei der Entstehung der Pellets spielen. Die vor allem aus Calcium und Fetts{\"a}uren bestehenden Pellets dienten als Aufwuchsfl{\"a}chen f{\"u}r verschiedene Mikroorganismen. Die Bildung von Biofilmen, wie sie auf und in den Pellets nachgewiesen wurde, bot f{\"u}r Mikroorganismen einen Schutz vor negativen Umwelteinfl{\"u}ssen wie z. B. hohe Propions{\"a}urekonzentrationen. Unter diesen g{\"u}nstigen Bedingungen war die Bildung von Biogas auch unter hohen Wasserstoffpartialdr{\"u}cken, die den Abbau von Propions{\"a}ure hemmten, m{\"o}glich. Als Indikator f{\"u}r bessere Lebensbedingungen wurde im Laborversuch ein Methanoculleus receptaculi-verwandter Organismus identifiziert. Dieses methanogene Archaeon wurde im Pellet nachgewiesen, w{\"a}hrend es im G{\"a}rrest erst nach der Prozessregenerierung detektiert wurde. Der Nachweis eines im Vergleich zum umgebenden G{\"a}rrest h{\"o}heren Anteils an Archaeen im Kern der Pellets sowie von Biofilmen/EPS, verschiedenen Phosphatsalzen und schwerl{\"o}slichen Calciumsalzen zeigte, dass sowohl Pr{\"a}zipitation und Adsorption als auch Degradation von LCFA dazu f{\"u}hren, dass deren Konzentration im fl{\"u}ssigen G{\"a}rrest gesenkt wird. Dadurch nimmt die Hemmung auf die Bioz{\"o}nose ab und die Biogasbildungsrate steigt. Daher ist der Abbau der Fetts{\"a}uren auch bei einem niedrigen pH-Wert und unter hohen Wasserstoffpartialdr{\"u}cken m{\"o}glich und der Biogasbildungsprozess ist langfristig stabil. Die Bildung von Pellets unterst{\"u}tzt die Prozessstabilit{\"a}t, sofern diese nicht zu groß werden und dann u. a. die Durchmischung behindern und den Ablauf verstopfen. Nach erfolgreicher Prozessstabilisierung wurden keine Pellets im G{\"a}rrest beobachtet. Der Abbau des organischen Materials wurde sowohl durch die steigende Calciumkonzentration als auch die steigende Gasproduktion angezeigt.}, language = {de} } @phdthesis{Olszewska2015, author = {Olszewska, Agata}, title = {Forming magnetic chain with the help of biological organisms}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-89767}, school = {Universit{\"a}t Potsdam}, pages = {101}, year = {2015}, abstract = {Magnetite nanoparticles and their assembly comprise a new area of development for new technologies. The magnetic particles can interact and assemble in chains or networks. Magnetotactic bacteria are one of the most interesting microorganisms, in which the assembly of nanoparticles occurs. These microorganisms are a heterogeneous group of gram negative prokaryotes, which all show the production of special magnetic organelles called magnetosomes, consisting of a magnetic nanoparticle, either magnetite (Fe3O4) or greigite (Fe3S4), embedded in a membrane. The chain is assembled along an actin-like scaffold made of MamK protein, which makes the magnetosomes to arrange in mechanically stable chains. The chains work as a compass needle in order to allow cells to orient and swim along the magnetic field of the Earth. The formation of magnetosomes is known to be controlled at the molecular level. The physico-chemical conditions of the surrounding environment also influence biomineralization. The work presented in this manuscript aims to understand how such external conditions, in particular the extracellular oxidation reduction potential (ORP) influence magnetite formation in the strain Magnetospirillum magneticum AMB-1. A controlled cultivation of the microorganism was developed in a bioreactor and the formation of magnetosomes was characterized. Different techniques have been applied in order to characterize the amount of iron taken up by the bacteria and in consequence the size of magnetosomes produced at different ORP conditions. By comparison of iron uptake, morphology of bacteria, size and amount of magnetosomes per cell at different ORP, the formation of magnetosomes was inhibited at ORP 0 mV, whereas reduced conditions, ORP - 500 mV facilitate biomineralization process. Self-assembly of magnetosomes occurring in magnetotactic bacteria became an inspiration to learn from nature and to construct nanoparticles assemblies by using the bacteriophage M13 as a template. The M13 bacteriophage is an 800 nm long filament with encapsulated single-stranded DNA that has been recently used as a scaffold for nanoparticle assembly. I constructed two types of assemblies based on bacteriophages and magnetic nanoparticles. A chain - like assembly was first formed where magnetite nanoparticles are attached along the phage filament. A sperm - like construct was also built with a magnetic head and a tail formed by phage filament. The controlled assembly of magnetite nanoparticles on the phage template was possible due to two different mechanism of nanoparticle assembly. The first one was based on the electrostatic interactions between positively charged polyethylenimine coated magnetite nanoparticles and negatively charged phages. The second phage -nanoparticle assembly was achieved by bioengineered recognition sites. A mCherry protein is displayed on the phage and is was used as a linker to a red binding nanobody (RBP) that is fused to the one of the proteins surrounding the magnetite crystal of a magnetosome. Both assemblies were actuated in water by an external magnetic field showing their swimming behavior and potentially enabling further usage of such structures for medical applications. The speed of the phage - nanoparticles assemblies are relatively slow when compared to those of microswimmers previously published. However, only the largest phage-magnetite assemblies could be imaged and it is therefore still unclear how fast these structures can be in their smaller version.}, language = {en} } @phdthesis{Lischke2015, author = {Lischke, Betty}, title = {Food web regulation under different forcing regimes in shallow lakes}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-89149}, school = {Universit{\"a}t Potsdam}, pages = {131}, year = {2015}, abstract = {The standing stock and production of organismal biomass depends strongly on the organisms' biotic environment, which arises from trophic and non-trophic interactions among them. The trophic interactions between the different groups of organisms form the food web of an ecosystem, with the autotrophic and bacterial production at the basis and potentially several levels of consumers on top of the producers. Feeding interactions can regulate communities either by severe grazing pressure or by shortage of resources or prey production, termed top-down and bottom-up control, respectively. The limitations of all communities conglomerate in the food web regulation, which is subject to abiotic and biotic forcing regimes arising from external and internal constraints. This dissertation presents the effects of alterations in two abiotic, external forcing regimes, terrestrial matter input and long-lasting low temperatures in winter. Diverse methodological approaches, a complex ecosystem model study and the analysis of two whole-lake measurements, were performed to investigate effects for the food web regulation and the resulting consequences at the species, community and ecosystem scale. Thus, all types of organisms, autotrophs and heterotrophs, at all trophic levels were investigated to gain a comprehensive overview of the effects of the two mentioned altered forcing regimes. In addition, an extensive evaluation of the trophic interactions and resulting carbon fluxes along the pelagic and benthic food web was performed to display the efficiencies of the trophic energy transfer within the food webs. All studies were conducted in shallow lakes, which is worldwide the most abundant type of lakes. The specific morphology of shallow lakes allows that the benthic production contributes substantially to the whole-lake production. Further, as shallow lakes are often small they are especially sensitive to both, changes in the input of terrestrial organic matter and the atmospheric temperature. Another characteristic of shallow lakes is their appearance in alternative stable states. They are either in a clear-water or turbid state, where macrophytes and phytoplankton dominate, respectively. Both states can stabilize themselves through various mechanisms. These two alternative states and stabilizing mechanisms are integrated in the complex ecosystem model PCLake, which was used to investigate the effects of the enhanced terrestrial particulate organic matter (t-POM) input to lakes. The food web regulation was altered by three distinct pathways: (1) Zoobenthos received more food, increased in biomass which favored benthivorous fish and those reduced the available light due to bioturbation. (2) Zooplankton substituted autochthonous organic matter in their diet by suspended t-POM, thus the autochthonous organic matter remaining in the water reduced its transparency. (3) T-POM suspended into the water and reduced directly the available light. As macrophytes are more light-sensitive than phytoplankton they suffered the most from the lower transparency. Consequently, the resilience of the clear-water state was reduced by enhanced t-POM inputs, which makes the turbid state more likely at a given nutrient concentration. In two subsequent winters long-lasting low temperatures and a concurrent long duration of ice coverage was observed which resulted in low overall adult fish biomasses in the two study lakes - Schulzensee and Gollinsee, characterized by having and not having submerged macrophytes, respectively. Before the partial winterkill of fish Schulzensee allowed for a higher proportion of piscivorous fish than Gollinsee. However, the partial winterkill of fish aligned both communities as piscivorous fish are more sensitive to low oxygen concentrations. Young of the year fish benefitted extremely from the absence of adult fish due to lower predation pressure. Therefore, they could exert a strong top-down control on crustaceans, which restructured the entire zooplankton community leading to low crustacean biomasses and a community composition characterized by copepodites and nauplii. As a result, ciliates were released from top-down control, increased to high biomasses compared to lakes of various trophic states and depths and dominated the zooplankton community. While being very abundant in the study lakes and having the highest weight specific grazing rates among the zooplankton, ciliates exerted potentially a strong top-down control on small phytoplankton and particle-attached bacteria. This resulted in a higher proportion of large phytoplankton compared to other lakes. Additionally, the phytoplankton community was evenly distributed presumably due to the numerous fast growing and highly specific ciliate grazers. Although, the pelagic food web was completely restructured after the subsequent partial winterkills of fish, both lakes were resistant to effects of this forcing regime at the ecosystem scale. The consistently high predation pressure on phytoplankton prevented that Schulzensee switched from the clear-water to the turbid state. Further mechanisms, which potentially stabilized the clear-water state, were allelopathic effects by macrophytes and nutrient limitation in summer. The pelagic autotrophic and bacterial production was an order of magnitude more efficient transferred to animal consumers than the respective benthic production, despite the alterations of the food web structure after the partial winterkill of fish. Thus, the compiled mass-balanced whole-lake food webs suggested that the benthic bacterial and autotrophic production, which exceeded those of the pelagic habitat, was not used by animal consumers. This holds even true if the food quality, additional consumers such as ciliates, benthic protozoa and meiobenthos, the pelagic-benthic link and the potential oxygen limitation of macrobenthos were considered. Therefore, low benthic efficiencies suggest that lakes are primarily pelagic systems at least at the animal consumer level. Overall, this dissertation gives insights into the regulation of organism groups in the pelagic and benthic habitat at each trophic level under two different forcing regimes and displays the efficiency of the carbon transfer in both habitats. The results underline that the alterations of external forcing regimes affect all hierarchical level including the ecosystem.}, language = {en} } @phdthesis{Borschewski2015, author = {Borschewski, Aljona}, title = {Bedeutung der Interaktion von Calcineurin und SORLA f{\"u}r die Regulation des Na⁺,K⁺,2Cl⁻-Kotransporters (NKCC2) in der Niere}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-89205}, school = {Universit{\"a}t Potsdam}, pages = {II, 86}, year = {2015}, abstract = {Der Na⁺-K⁺-2Cl⁻-Kotransporter (NKCC2) wird im distalen Nephron der Niere exprimiert. Seine Verteilung umfasst die Epithelien der medull{\"a}ren und kortikalen Teile der dicken aufsteigenden Henle-Schleife (Thick ascending limb, TAL) und die Macula densa. Resorptiver NaCl-Transport {\"u}ber den NKCC2 dient dem renalen Konzentrierungsmechanismus und reguliert systemisch auch Volumenstatus und Blutdruck. Die Aktivit{\"a}t des NKCC2 ist mit der Phosphorylierung seiner N-terminalen Aminos{\"a}urereste Serin 126 und Threonin 96/101 verbunden. Vermittelt wird diese durch die homologen Kinasen SPAK (SPS-related proline/alanine-rich kinase) und OSR1 (Oxidative stress responsive kinase 1), die hierzu ihrerseits phosphoryliert werden m{\"u}ssen. Der regulatorische Kontext dieser Kinasen ist mittlerweile gut charakterisiert. {\"U}ber Mechanismen und Produkte, die den NKCC2 deaktivieren, war hingegen weniger bekannt. Ziel der Arbeit war daher zu untersuchen, welche Wege zur Deaktivierung des Transporters f{\"u}hren. Der intrazellul{\"a}re Sortierungsrezeptor SORLA (Sorting-protein-related receptor with A-type repeats) war zuvor in seiner Bedeutung f{\"u}r das Nephron charakterisiert worden. Ein SORLA-defizientes Mausmodell weist unter anderem eine stark verringerte NKCC2-Phosphorylierung auf. Unter osmotischem Stress k{\"o}nnen SORLA-defiziente M{\"a}use ihren Urin weniger effizient konzentrieren. Meine Resultate zeigen mit hochaufl{\"o}sender Technik, dass SORLA apikal im TAL lokalisiert ist und dass mit NKCC2 eine anteilige Kolokalisation besteht. Unter SORLA Defizienz war die f{\"u}r die NKCC2 Aktivit{\"a}t maßgebliche SPAK/OSR1-Phosphorylierung gegen{\"u}ber dem Wildtyp nicht ver{\"a}ndert. Jedoch war die ebenfalls im TAL exprimierte Phosphatase Calcineurin Aβ (CnAβ) per Western blot um das zweifache gesteigert. Parallel hierzu wurde immunhistochemisch die Kolokalisation von verst{\"a}rktem CnAβ-Signal und NKCC2 best{\"a}tigt. Beide Befunde geben zusammen den Hinweis auf einen Bezug zwischen der reduzierten NKCC2-Phosphorylierung und der gesteigerten Pr{\"a}senz von CnAβ bei SORLA Defizienz. Die parallel induzierte {\"U}berexpression von SORLA in HEK-Zellen zeigte entsprechend eine Halbierung der CnAβ Proteinmenge. SORLA steuert demzufolge sowohl die Abundanz als auch die zellul{\"a}re Verteilung der Phosphatase. Weiterhin ließ sich die Interaktion zwischen CnAβ und SORLA (intrazellul{\"a}re Dom{\"a}ne) mittels Co-Immunpr{\"a}zipitation bzw. GST-pulldown assay nachweisen. Auch die Interaktion zwischen CnAβ und NKCC2 wurde auf diesem Weg belegt. Da allerdings weder SORLA noch NKCC2 ein spezifisches Bindungsmuster f{\"u}r CnAβ aufweisen, sind vermutlich intermedi{\"a}re Adapterproteine bei ihrer Bindung involviert. Die pharmakologische Inhibition von CnAβ mittels Cyclosporin A (CsA; 1 h) f{\"u}hrte bei SORLA Defizienz zur Normalisierung der NKCC2-Phosphorylierung. Entsprechend f{\"u}hrte in vitro die Gabe von CsA bei TAL Zellen zu einer 7-fach gesteigerten NKCC2-Phosphorylierung. Zusammenfassend zeigen die Ergebnisse, dass die Phosphatase CnAβ {\"u}ber ihre Assoziation mit NKCC2 diesen im adluminalen Zellkompartiment deaktivieren kann. Gesteuert wird dieser Vorgang durch die Eigenschaft von SORLA, CnAβ apikal zu reduzieren und damit die adluminale Phosphorylierung und Aktivit{\"a}t von NKCC2 zu unterst{\"u}tzen. Da Calcineurin-Inhibitoren derzeit die Grundlage der immunsupprimierenden Therapie darstellen, haben die Ergebnisse eine klinische Relevanz. Angesichts der Co-Expression von SORLA und CnAβ in verschiedenen anderen Organen k{\"o}nnen die Ergebnisse auch {\"u}ber die Niere hinaus Bedeutung erlangen.}, language = {de} } @misc{LamannaKirschbaumWauricketal.2015, author = {Lamanna, Francesco and Kirschbaum, Frank and Waurick, Isabelle and Dieterich, Christoph and Tiedemann, Ralph}, title = {Cross-tissue and cross-species analysis of gene expression in skeletal muscle and electric organ of African weakly-electric fish (Teleostei; Mormyridae)}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-86997}, year = {2015}, abstract = {Background African weakly-electric fishes of the family Mormyridae are able to produce and perceive weak electric signals (typically less than one volt in amplitude) owing to the presence of a specialized, muscle-derived electric organ (EO) in their tail region. Such electric signals, also known as Electric Organ Discharges (EODs), are used for objects/prey localization, for the identification of conspecifics, and in social and reproductive behaviour. This feature might have promoted the adaptive radiation of this family by acting as an effective pre-zygotic isolation mechanism. Despite the physiological and evolutionary importance of this trait, the investigation of the genetic basis of its function and modification has so far remained limited. In this study, we aim at: i) identifying constitutive differences in terms of gene expression between electric organ and skeletal muscle (SM) in two mormyrid species of the genus Campylomormyrus: C. compressirostris and C. tshokwe, and ii) exploring cross-specific patterns of gene expression within the two tissues among C. compressirostris, C. tshokwe, and the outgroup species Gnathonemus petersii. Results Twelve paired-end (100 bp) strand-specific RNA-seq Illumina libraries were sequenced, producing circa 330 M quality-filtered short read pairs. The obtained reads were assembled de novo into four reference transcriptomes. In silico cross-tissue DE-analysis allowed us to identify 271 shared differentially expressed genes between EO and SM in C. compressirostris and C.tshokwe. Many of these genes correspond to myogenic factors, ion channels and pumps, and genes involved in several metabolic pathways. Cross-species analysis has revealed that the electric organ transcriptome is more variable in terms of gene expression levels across species than the skeletal muscle transcriptome. Conclusions The data obtained indicate that: i) the loss of contractile activity and the decoupling of the excitation-contraction processes are reflected by the down-regulation of the corresponding genes in the electric organ's transcriptome; ii) the metabolic activity of the EO might be specialized towards the production and turn-over of membrane structures; iii) several ion channels are highly expressed in the EO in order to increase excitability; iv) several myogenic factors might be down-regulated by transcription repressors in the EO.}, language = {en} } @phdthesis{Lamanna2015, author = {Lamanna, Francesco}, title = {Adaptive radiation and speciation in African weakly-electric fish}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-80097}, school = {Universit{\"a}t Potsdam}, pages = {114}, year = {2015}, abstract = {The rise of evolutionary novelties is one of the major drivers of evolutionary diversification. African weakly-electric fishes (Teleostei, Mormyridae) have undergone an outstanding adaptive radiation, putatively owing to their ability to communicate through species-specific Electric Organ Discharges (EODs) produced by a novel, muscle-derived electric organ. Indeed, such EODs might have acted as effective pre-zygotic isolation mechanisms, hence favoring ecological speciation in this group of fishes. Despite the evolutionary importance of this organ, genetic investigations regarding its origin and function have remained limited. The ultimate aim of this study is to better understand the genetic basis of EOD production by exploring the transcriptomic profiles of the electric organ and of its ancestral counterpart, the skeletal muscle, in the genus Campylomormyrus. After having established a set of reference transcriptomes using "Next-Generation Sequencing" (NGS) technologies, I performed in silico analyses of differential expression, in order to identify sets of genes that might be responsible for the functional differences observed between these two kinds of tissues. The results of such analyses indicate that: i) the loss of contractile activity and the decoupling of the excitation-contraction processes are reflected by the down-regulation of the corresponding genes in the electric organ; ii) the metabolic activity of the electric organ might be specialized towards the production and turnover of membrane structures; iii) several ion channels are highly expressed in the electric organ in order to increase excitability, and iv) several myogenic factors might be down-regulated by transcription repressors in the EO. A secondary task of this study is to improve the genus level phylogeny of Campylomormyrus by applying new methods of inference based on the multispecies coalescent model, in order to reduce the conflict among gene trees and to reconstruct a phylogenetic tree as closest as possible to the actual species-tree. By using 1 mitochondrial and 4 nuclear markers, I was able to resolve the phylogenetic relationships among most of the currently described Campylomormyrus species. Additionally, I applied several coalescent-based species delimitation methods, in order to test the hypothesis that putatively cryptic species, which are distinguishable only from their EOD, belong to independently evolving lineages. The results of this analysis were additionally validated by investigating patterns of diversification at 16 microsatellite loci. The results suggest the presence of a new, yet undescribed species of Campylomormyrus.}, language = {en} } @misc{SbragagliaLamannaMatetal.2015, author = {Sbragaglia, Valerio and Lamanna, Francesco and Mat, Audrey M. and Rotllant, Guiomar and Joly, Silvia and Ketmaier, Valerio and de la Iglesia, Horacio O. and Aguzzi, Jacopo}, title = {Identification, Characterization, and Diel Pattern of Expression of Canonical Clock Genes in Nephrops norvegicus (Crustacea: Decapoda) Eyestalk}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-84432}, year = {2015}, abstract = {The Norway lobster, Nephrops norvegicus, is a burrowing decapod with a rhythmic burrow emergence (24 h) governed by the circadian system. It is an important resource for European fisheries and its behavior deeply affects its availability. The current knowledge of Nephrops circadian biology is phenomenological as it is currently the case for almost all crustaceans. In attempt to elucidate the putative molecular mechanisms underlying circadian gene regulation in Nephrops, we used a transcriptomics approach on cDNA extracted from the eyestalk, a structure playing a crucial role in controlling behavior of decapods. We studied 14 male lobsters under 12-12 light-darkness blue light cycle. We used the Hiseq 2000 Illumina platform to sequence two eyestalk libraries (under light and darkness conditions) obtaining about 90 millions 100-bp paired-end reads. Trinity was used for the de novo reconstruction of transcriptomes; the size at which half of all assembled bases reside in contigs (N50) was equal to 1796 (light) and 2055 (darkness). We found a list of candidate clock genes and focused our attention on canonical ones: timeless, period, clock and bmal1. The cloning of assembled fragments validated Trinity outputs. The putative Nephrops clock genes showed high levels of identity (blastx on NCBI) with known crustacean clock gene homologs such as Eurydice pulchra (period: 47\%, timeless: 59\%, bmal1: 79\%) and Macrobrachium rosenbergii (clock: 100\%). We also found a vertebrate-like cryptochrome 2. RT-qPCR showed that only timeless had a robust diel pattern of expression. Our data are in accordance with the current knowledge of the crustacean circadian clock, reinforcing the idea that the molecular clockwork of this group shows some differences with the established model in Drosophila melanogaster.}, language = {en} } @phdthesis{Reim2015, author = {Reim, Tina}, title = {Biogene Aminrezeptoren bei der Honigbiene Apis mellifera}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-80982}, school = {Universit{\"a}t Potsdam}, pages = {viii, 106}, year = {2015}, abstract = {Die Honigbiene Apis mellifera zeigt innerhalb einer Kolonie eine an das Alter gekoppelte Arbeitsteilung. Junge Honigbienen versorgen die Brut (Ammenbienen), w{\"a}hrend {\"a}ltere Honigbienen (Sammlerinnen) außerhalb des Stocks Pollen und Nektar eintragen. Die biogenen Amine Octopamin und Tyramin sind an der Steuerung der Arbeitsteilung maßgeblich beteiligt. Sie interagieren mit Zielzellen {\"u}ber die Bindung an G Protein gekoppelte Rezeptoren. A. mellifera besitzt f{\"u}nf charakterisierte Octopaminrezeptoren (AmOctαR1, AmOctβR1-4), einen charakterisierten Tyraminrezeptor (AmTyr1) sowie einen weiteren putativen Tyraminrezeptor. In der vorliegenden Arbeit wurde dieser putative Aminrezeptor als zweiter Tyraminrezeptor (AmTyr2) identifiziert, lokalisiert und pharmakologisch charakterisiert. Die von der cDNA abgeleitete Aminos{\"a}uresequenz weist strukturelle Eigenschaften und konservierte Motive von G Protein gekoppelten Rezeptoren auf. Phylogenetisch ordnet sich der AmTyr2 Rezeptor bei den Tyramin 2 Rezeptoren anderer Insekten ein. Die funktionelle und pharmakologische Charakterisierung des putativen Tyraminrezeptors erfolgte in modifizierten HEK293 Zellen, die mit der Rezeptor cDNA transfiziert wurden. Die Applikation von Tyramin aktiviert Adenylylcyclasen in diesen Zellen und resultiert in einem Anstieg des intrazellul{\"a}ren cAMP Gehalts. Der AmTyr2 Rezeptor kann durch Tyramin in nanomolaren Konzentrationen halbmaximal aktiviert werden. W{\"a}hrend es sich bei Octopamin um einen wirkungsvollen Agonisten des Rezeptors handelt, sind Mianserin und Yohimbin effektive Antagonisten. F{\"u}r die Lokalisierung des Rezeptorproteins wurde ein polyklonaler Antik{\"o}rper generiert. Eine AmTyr2-{\"a}hnliche Immunreaktivit{\"a}t zeigt sich im Gehirn in den optischen Loben, den Antennalloben, dem Zentralkomplex und in den Kenyon Zellen der Pilzk{\"o}rper. Des Weiteren wurde die Rolle der Octopamin- und Tyraminrezeptoren bei der Steuerung der altersabh{\"a}ngigen Arbeitsteilung analysiert. Die Genexpression des AmOctαR1 in verschiedenen Gehirnteilen korreliert unabh{\"a}ngig vom Alter mit der sozialen Rolle, w{\"a}hrend sich die Genexpression von AmOctβR3/4 und den Tyraminrezeptoren AmTyr1 und AmTyr2 maximal mit dem Alter aber nicht der sozialen Rolle {\"a}ndert. Sammlerinnen weisen einen h{\"o}heren Octopamingehalt im Gesamtgehirn auf als Ammenbienen; bei Tyramin zeigen sich keine Unterschiede. W{\"a}hrend Tyramin offensichtlich keine direkte Rolle spielt, werden durch Octopamin gesteuerte Prozesse der altersabh{\"a}ngigen Arbeitsteilung bei der Honigbiene vermutlich {\"u}ber den AmOctαR1 vermittelt. Die Ergebnisse der vorliegenden Arbeit zeigen die wichtige Rolle von biogenen Aminen, insbesondere Octopamin bei der sozialen Organisation von Insektenstaaten.}, language = {de} } @phdthesis{Schmidt2015, author = {Schmidt, Andreas}, title = {Charakterisierung der Lipopolysaccharid-Bindungseigenschaften von Adh{\"a}sionsproteinen aus Salmonella-Bakteriophagen}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-79529}, school = {Universit{\"a}t Potsdam}, pages = {VIII, 114}, year = {2015}, abstract = {Die Interaktionen von komplexen Kohlenhydraten und Proteinen sind ubiquit{\"a}r. Sie spielen wichtige Rollen in vielen physiologischen Prozessen wie Zelladh{\"a}sion, Signaltransduktion sowie bei viralen Infektionen. Die molekularen Grundlagen der Interaktion sind noch nicht komplett verstanden. Ein Modellsystem f{\"u}r Kohlenhydrat-Protein-Interaktionen besteht aus Adh{\"a}sionsproteinen (Tailspikes) von Bakteriophagen, die komplexe Kohlenhydrate auf bakteriellen Oberfl{\"a}chen (O-Antigen) erkennen. Das Tailspike-Protein (TSP), das in dieser Arbeit betrachtet wurde, stammt aus dem Bakteriophagen 9NA (9NATSP). 9NATSP weist eine hohe strukturelle Homologie zum gut charakterisierten TSP des Phagen P22 (P22TSP) auf, bei einer niedriger sequenzieller {\"A}hnlichkeit. Die Substratspezifit{\"a}ten beider Tailspikes sind {\"a}hnlich mit Ausnahme der Toleranz gegen{\"u}ber den glucosylierten Formen des O-Antigens. Die Struktur der beiden Tailspikes ist bekannt, sodass sie ein geeignetes System f{\"u}r vergleichende Bindungsstudien darstellen, um die strukturellen Grundlagen f{\"u}r die Unterschiede der Spezifit{\"a}t zu untersuchen. Im Rahmen dieser Arbeit wurde der ELISA-like tailspike adsorption assay (ELITA) etabliert, um Binderpaare aus TSPs und O-Antigen zu identifizieren. Dabei wurden 9NATSP und P22TSP als Sonden eingesetzt, deren Bindung an die intakten, an die Mikrotiterplatte adsorbierten Bakterien getestet wurde. Beim Test einer Sammlung aus 44 Salmonella-St{\"a}mmen wurden St{\"a}mme identifiziert, die bindendes O-Antigen exprimieren. Gleichzeitig wurden Unterschiede in der Bindung der beiden TSPs an Salmonella-St{\"a}mme mit gleichem O-Serotyp beobachtet. Die Ergebnisse der ELITA-Messung wurden qualitativ durch eine FACS-basierte Bindungsmessung best{\"a}tigt. Zus{\"a}tzlich erm{\"o}glichte die FACS-Messung bei St{\"a}mmen, die teilweise modifizierte O-Antigene herstellen, den Anteil an Zellen mit und ohne Modifikation zu erfassen. Die Oberfl{\"a}chenplasmonresonanz (SPR)-basierten Interaktionsmessungen wurden eingesetzt, um Bindungsaffinit{\"a}ten f{\"u}r eine TSP-O-Antigen Kombination zu quantifizieren. Daf{\"u}r wurden zwei Methoden getestet, um die Oligosaccharide auf einem SPR-Chip zu immobilisieren. Zum einen wurden die enzymatisch hergestellten O-Antigenfragmente mit einem bifunktionalen Oxaminadapter derivatisiert, der eine prim{\"a}re Aminogruppe f{\"u}r die Immobilisierung bereitstellt. Ein Versuch, diese Oligosaccharidfragmente zu immobilisieren, war jedoch nicht erfolgreich. Dagegen wurde das nicht derivatisierte Polysaccharid, bestehend aus repetitivem O-Antigen und einem konservierten Kernsaccharid, erfolgreich auf einem SPR-Chip immobilisiert. Die Immobilisierung wurde durch Interaktionsmessungen mit P22TSP best{\"a}tigt. Durch die Immobilisierung des Polysaccharids sind somit quantitative SPR-Bindungsmessungen mit einem polydispersen Interaktionspartner m{\"o}glich. Eine Auswahl von Salmonella-St{\"a}mmen mit einer ausgepr{\"a}gt unterschiedlichen Bindung von 9NATSP und P22TSP im ELITA-Testsystem wurde hinsichtlich der Zusammensetzung des O-Antigens mittels HPLC, Kapillargelelektrophorese und MALDI-MS analysiert. Dabei wurden nicht-st{\"o}chiometrische Modifikationen der O-Antigene wie Acetylierung und Glucosylierung detektiert. Das Ausmaß der Glucosylierung korrelierte negativ mit der Effizienz der Bindung und des Verdaus durch die beiden TSPs, wobei der negative Effekt bei 9NATSP weniger stark ausgepr{\"a}gt war als bei P22TSP. Dies stimmt mit den Literaturdaten zu Infektivit{\"a}tsstudien mit 9NA und P22 {\"u}berein, die mit St{\"a}mmen mit vergleichbaren O-Antigenvarianten durchgef{\"u}hrt wurden. Die Korrelation zwischen der Glucosylierung und Bindungseffizienz konnte strukturell interpretiert werden. Auf Grundlage der O-Antigenanalysen sowie der Ergebnisse der ELITA- und FACS-Bindungstests wurden die Salmonella-St{\"a}mme Brancaster und Kalamu identifiziert, die ann{\"a}hernd quantitativ glucosyliertes O-Antigen exprimieren. Damit eignen sich diese St{\"a}mme f{\"u}r weiterf{\"u}hrende Studien, um die Zusammenh{\"a}nge zwischen der Spezifit{\"a}t und der Organisation der Bindestellen der beiden TSPs zu untersuchen.}, language = {de} } @phdthesis{Wettstein2015, author = {Wettstein, Christoph}, title = {Cytochrome c-DNA and cytochrome c-enzyme interactions for the construction of analytical signal chains}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-78367}, school = {Universit{\"a}t Potsdam}, pages = {120}, year = {2015}, abstract = {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.}, language = {en} } @phdthesis{Niedl2015, author = {Niedl, Robert Raimund}, title = {Nichtlineare Kinetik und responsive Hydrogele f{\"u}r papierbasierte Schnelltestanwendungen}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-77735}, school = {Universit{\"a}t Potsdam}, pages = {iv, 128}, year = {2015}, abstract = {Viele klinische Schnelltestsysteme ben{\"o}tigen vorpr{\"a}parierte oder aufgereinigte Analyte mit frisch hergestellten L{\"o}sungen. Fernab standardisierter Laborbedingungen wie z.B. in Entwicklungsl{\"a}ndern oder Krisengebieten sind solche Voraussetzungen oft nur unter einem hohen Aufwand herstellbar. Zus{\"a}tzlich stellt die erforderliche Sensitivit{\"a}t die Entwicklung einfach zu handhabender Testsysteme vor große Herausforderungen. Autokatalytische Reaktionen, die sich mit Hilfe sehr geringer Initiatorkonzentrationen ausl{\"o}sen lassen, k{\"o}nnen hier eine Perspektive f{\"u}r Signalverst{\"a}rkungsprozesse bieten. Aus diesem Grund wird im ersten Teil der vorliegenden Arbeit das Verhalten der autokatalytischen Arsenit-Jodat-Reaktion in einem mikrofluidischen Kanal untersucht. Dabei werden insbesondere die diffusiven und konvektiven Einfl{\"u}sse auf die Reaktionskinetik im Vergleich zu makroskopischen Volumenmengen betrachtet. Im zweiten Teil werden thermoresponsive Hydrogele mit einem kanalstrukturierten Papiernetzwerk zu einem neuartigen, kapillargetriebenen, extern steuerbaren Mikrofluidik-System kombiniert. Das hier vorgestellte Konzept durch Hydrogele ein papierbasiertes LOC-System zu steuern, erm{\"o}glicht zuk{\"u}nftig die Herstellung von komplexeren, steuerbaren Point-Of-Care Testsystemen (POCT). Durch z.B. einen thermischen Stimulus, wird das L{\"o}sungsverhalten eines Hydrogels so ver{\"a}ndert, dass die gespeicherte Fl{\"u}ssigkeit freigesetzt und durch die Kapillarkraft des Papierkanals ins System transportiert wird. Die Eigenschaften dieses Gelnetzwerks k{\"o}nnen dabei so eingestellt werden, dass eine Freisetzung von Fl{\"u}ssigkeiten sogar bei K{\"o}rpertemperatur m{\"o}glich w{\"a}re und damit eine Anwendung g{\"a}nzlich ohne weitere Hilfsmittel denkbar ist. F{\"u}r die Anwendung notwendige Chemikalien oder Enzyme lassen sich hierbei bequem in getrocknetem Zustand im Papiersubstrat vorlagern und bei Bedarf in L{\"o}sung bringen. Im abschließenden dritten Teil der Arbeit wird ein durch Hydrogele betriebener, Antik{\"o}rper-basierter Mikroorganismenschnelltest f{\"u}r Escherichia coli pr{\"a}sentiert. Dar{\"u}ber hinaus wird weiterf{\"u}hrend eine einfache Methode zur Funktionalisierung eines Hydrogels mit Biomolek{\"u}len {\"u}ber EDC/NHS-Kopplung vorgestellt.}, language = {de} } @phdthesis{Rajasundaram2015, author = {Rajasundaram, Dhivyaa}, title = {Integrative analysis of heterogeneous plant cell wall related data}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-77652}, school = {Universit{\"a}t Potsdam}, pages = {xii, 205}, year = {2015}, abstract = {Plant cell walls are complex structures that underpin plant growth and are widely exploited in diverse human activities thus placing them with a central importance in biology. Cell walls have been a prominent area of research for a long time, but the chemical complexity and diversity of cell walls not just between species, but also within plants, between cell-types, and between cell wall micro-domains pose several challenges. Progress accelerated several-fold in cell wall biology owing to advances in sequencing technology, aided soon thereafter by advances in omics and imaging technologies. This development provides additional perspectives of cell walls across a rapidly growing number of species, highlighting a myriad of architectures, compositions, and functions. Furthermore, rather than the component centric view, integrative analysis of the different cell wall components across system-levels help to gain a more in-depth understanding of the structure and biosynthesis of the cell envelope and its interactions with the environment. To this end, in this work three case studies are detailed, all pertaining to the integrative analysis of heterogeneous cell wall related data arising from different system-levels and analytical techniques. A detailed account of multiblock methods is provided and in particular canonical correlation and regression methods of data integration are discussed. In the first integrative analysis, by employing canonical correlation analysis - a multivariate statistical technique to study the association between two datasets - novel insight to the relationship between glycans and phenotypic traits is gained. In addition, sparse partial least squares regression approach that adapts Lasso penalization and allows for the selection of a subset of variables was employed. The second case study focuses on an integrative analysis of images obtained from different spectroscopic techniques. By employing yet another multiblock approach - multiple co-inertia analysis, insitu biochemical composition of cell walls from different cell-types is studied thereby highlighting the common and complementary parts of the two hyperspectral imaging techniques. Finally, the third integrative analysis facilitates gene expression analysis of the Arabidopsis root transcriptome and translatome for the identification of cell wall related genes and compare expression patterns of cell wall synthesis genes. The computational analysis considered correlation and variation of expression across cell-types at both system-levels, and also provides insight into the degree of co-regulatory relationships that are preserved between the two processes. The integrative analysis of glycan data and phenotypic traits in cotton fibers using canonical methods led to the identification of specific polysaccharides which may play a major role during fiber development for the final fiber characteristics. Furthermore, this analysis provides a base for future studies on glycan arrays in case of developing cotton fibers. The integrative analysis of images from infrared and Raman spectroscopic approaches allowed the coupling of different analytical techniques to characterize complex biological material, thereby, representing various facets of their chemical properties. Moreover, the results from the co-inertia analysis demonstrated that the study was well adapted as it is relevant for coupling data tables in a symmetric way. Several indicators are proposed to investigate how the global and block scores are related. In addition, studying the root cells of \textit{Arabidopsis thaliana} allowed positing a novel pipeline to systematically investigate and integrate the different levels of information available at the global and single-cell level. The conducted analysis also confirms that previously identified key transcriptional activators of secondary cell wall development display highly conserved patterns of transcription and translation across the investigated cell-types. Moreover, the biological processes that display conserved and divergent patterns based on the cell-type-specific expression and translation levels are identified.}, language = {en} } @phdthesis{Schirmack2015, author = {Schirmack, Janosch}, title = {Activity of methanogenic archaea under simulated Mars analog conditions}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-73010}, school = {Universit{\"a}t Potsdam}, pages = {VI, 108}, year = {2015}, abstract = {Assumed comparable environmental conditions of early Mars and early Earth in 3.7 Ga ago - at a time when first fossil records of life on Earth could be found - suggest the possibility of life emerging on both planets in parallel. As conditions changed, the hypothetical life on Mars either became extinct or was able to adapt and might still exist in biological niches. The controversial discussed detection of methane on Mars led to the assumption, that it must have a recent origin - either abiotic through active volcanism or chemical processes, or through biogenic production. Spatial and seasonal variations in the detected methane concentrations and correlations between the presence of water vapor and geological features such as subsurface hydrogen, which are occurring together with locally increased detected concentrations of methane, gave fuel to the hypothesis of a possible biological source of the methane on Mars. Therefore the phylogenetically old methanogenic archaea, which have evolved under early Earth conditions, are often used as model-organisms in astrobiological studies to investigate the potential of life to exist in possible extraterrestrial habitats on our neighboring planet. In this thesis methanogenic archaea originating from two extreme environments on Earth were investigated to test their ability to be active under simulated Mars analog conditions. These extreme environments - the Siberian permafrost-affected soil and the chemoautotrophically based terrestrial ecosystem of Movile cave, Romania - are regarded as analogs for possible Martian (subsurface) habitats. Two novel species of methanogenic archaea isolated from these environments were described within the frame of this thesis. It could be shown that concentrations up to 1 wt\% of Mars regolith analogs added to the growth media had a positive influence on the methane production rates of the tested methanogenic archaea, whereas higher concentrations resulted in decreasing rates. Nevertheless it was possible for the organisms to metabolize when incubated on water-saturated soil matrixes made of Mars regolith analogs without any additional nutrients. Long-term desiccation resistance of more than 400 days was proven with reincubation and indirect counting of viable cells through a combined treatment with propidium monoazide (to inactivate DNA of destroyed cells) and quantitative PCR. Phyllosilicate rich regolith analogs seem to be the best soil mixtures for the tested methanogenic archaea to be active under Mars analog conditions. Furthermore, in a simulation chamber experiment the activity of the permafrost methanogen strain Methanosarcina soligelidi SMA-21 under Mars subsurface analog conditions could be proven. Through real-time wavelength modulation spectroscopy measurements the increase in the methane concentration at temperatures down to -5 °C could be detected. The results presented in this thesis contribute to the understanding of the activity potential of methanogenic archaea under Mars analog conditions and therefore provide insights to the possible habitability of present-day Mars (near) subsurface environments. Thus, it contributes also to the data interpretation of future life detection missions on that planet. For example the ExoMars mission of the European Space Agency (ESA) and Roscosmos which is planned to be launched in 2018 and is aiming to drill in the Martian subsurface.}, language = {en} }