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Neuroligine, synaptische Regulation und Sozialverhalten von Mensch und Fliege

 (S. 476)
Nina Hahn, Ralf Heinrich

Neuroligins represent a family of phylogenetically conserved adhesion molecules that regulate synaptic physiology. Their malfunction is asso - ciated with autism spectrum disorders. Courtship and aggression of Drosophila melanogaster can serve as a model system to study the function of neuroligins in the proper execution of social behaviours, communication and cognitive tasks, the functions that are compromised in autistic patients.

Bakterielle Megastädte

 (S. 480)
Regine Hengge

What makes a multicellular aggregate of bacterial cells a biofilm is the presence of an extracellular polymeric matrix. Besides being protective, matrix components such as amyloid curli fibres and cellulose in Escherichia coli<(i> also assemble into a complex architecture that is controlled by RpoS and c-di-GMP. This confers extreme cohesion and elasticity to macrocolony biofilms. As a consequence, these biofilms behave like tissues, i. e. they buckle up into intricate morphological structures during growth.

Zelluläre Logistik der Mitochondrienvererbung

 (S. 484)
Stefan Böckler, Nadine Hock, Benedikt Westermann

Partitioning and inheritance of mitochondria upon cell division depends on cytoskeleton-dependent transport and organelle anchoring sites at the cell cortex. In asymmetrically dividing cells the mitochondria of both daughter cells may differ in their properties. In yeast cells intact mitochondria are transported towards the bud whereas old and damaged organelles are retained in the mother cell. Similarly, in stem cell division young mitochondria are selected to maintain stem cell properties.

Enzymatische Ligninverwertung durch bakterielle β-Etherasen

 (S. 487)
Anett Schallmey

Glutathione-dependent β-etherases are emerging enzymes for the bio - catalytic depolymerization of lignin, a heterogeneous aromatic polymer abundant in nature. They selectively catalyze the reductive cleavage of β-O-4 arylether bonds which account for 45 to 60 percent of linkages present in lignin. Hence, their application in lignin depolymerisation could enable a specific lignin breakdown, selectively yielding (valuable) lowmolecular- mass aromatics.

Systembiologie, Toxine und Epidemiologie von Clostridium difficile

 (S. 490)
Dieter Jahn

Clostridium difficile is increasingly causing deadly infections. It is growing in the gut and produces toxins due to microbiome deficiency caused by antibiotic treatment. Stool transplants are currently the only efficient strategy for the treatment of recurrent infections. An obvious lack of basic knowledge due to the difficult cultivation and genetic manipulation of these bacteria is a major challenge. The recently founded CDiff consortium is now investigating its systems biology, infection, and epidemiology.

Neue Einsichten in die Funktionsweise der inneren Uhr

 (S. 493)
Torsten Merbitz-Zahradnik, Eva Wolf

Most organisms exhibit daily rhythms in physiology and behaviour. In humans, the brain’s master clock is directly synchronized to the environmental light-dark cycle, while peripheral body clocks are set by master clock signals and the cell’s metabolic state. Structural analyses of the mammalian clock protein Cryptochrome (CRY) and its complex with the clock protein Period (PER) shows, how the inner clock could sense the cell’s metabolic state via CRY and its zinc-dependent interaction with PER.

Proteomik-Werkzeuge für Mikroorganismen

 (S. 496)
Ralf Rabus

Microorganisms play central roles in global elemental cycles and eco - system functioning. The molecular basis and mechanisms of the various ecophysiological capacities of environmental microbes are, however, often unknown or not clearly understood. In this regard, proteomic approaches promise plenty of opportunities. This short note recapitulates state-of-the-art proteomic tools for environmental microbiology and presents selected research examples.

Molekulare Identifikation von Hantaviren in neuen Wirten

 (S. 503)
Peter T. Witkowski, Patrick Heinemann, Boris Klempa, Detlev H. Krüger

In addition to classical virus isolation in cell culture, the molecular detection of new virus variants by PCR techniques allows broader epidemiological insights into the world of viral pathogens. For the detection of hantaviruses – zoonotic viruses leading to fever and organ failure in humans – we developed a genus-wide nested RT-PCR format, which enables the discovery of new members within this virus genus. The methodological approach allowed the demonstration of first hantaviruses from Africa and revealed new hantavirus reservoir hosts, as shrews, moles, and bats.

Digitale PCR in der Labordiagnostik

 (S. 507)
Gordon Handt, Mario Menschikowski, Werner Lehmann, Peter Schierack, Stefan Rödiger

The need for new highly sensitive, cost-efficient, fast and robust nucleic acid detection and quantification technologies is a driving force. PCR, especially quantitative PCR (qPCR), is the method of choice in diagnostics and life-sciences. The digital PCR (dPCR) provides a new technology to measure absolute quantities of nucleic acids without the need for calibration curves. This review gives details on the dPCR technology and available platforms. We discuss the platform differences, common features as well as their advantages and disadvantages.

Duplex-Real-Time-PCR zur Zygositätsbestimmung in transgenen Pflanzen

 (S. 511)
Leonie Davina Fritsch, Stefan Schillberg, Florian Schröper

Reliable techniques to detect the presence and zygosity of transgenic events in plants are crucial for biotechnology research and plant breeding industry. Therefore, we have introduced a novel assay, which enables high-throughput genotyping of transgenic events in maize using duplex real-time PCR. We demonstrated that the presence and zygosity of two selected transgenic events could be detected with high reproducibility and that our method is suitable for rapid genotyping of a large number of samples.

Anwendungen & Produkte

Automatisierte Herstellung von ChIP-Seq-Sequenzbibliotheken

 (S. 526)
Ulrike Bönisch, Laura Arrigoni, Emily Betancourt, Kerstin Bruder

Manual library preparation for ChIP-Seq applications is very tedious to conduct and lacks process standardization. Here we translate a commercially available ChIP-Seq library preparation protocol into commands for the Eppendorf epMotion automated liquid handling system and show that libraries with comparable quantity and quality can be easily generated, liberating lab personnel from repetitive processing steps while maximi - zing reproducibility and data comparability.


Analyse intrazellulärer Stoffflüsse in Säugerzellen

 (S. 564)
Sabine Herbst, Björn H. Junker

Analysis of metabolic fluxes enables a detailed view on effects of various genetic and environmental factors on the intracellular metabolism of different cell types. In mammalian cell culture metabolic flux analysis is mainly used to optimize cultivation processes for the production of biopharmaceuticals or to investigate the mode of action and toxicity of potential new drugs in medical research. Here, we give an overview about the workflow of metabolic flux analysis and areas of its application.



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Aktuell: PCR, qPCR

Aktuell: PCR, qPCR

Die Taq-Polymerase ist die zentrale Komponente der Polymerase-Kettenreaktion (polymerase chain reaction, PCR). Thomas D. Brock und Hudson Freeze von der Indiana University entdeckten 1969 diese thermostabile DNA-Polymerase in dem Bakterium Thermus aquaticus in einer heißen Quelle im Yellowstone-Nationalpark. Die Idee zur PCR wurde erstmals in den 1970er Jahren von Kjell Kleppe et al., Massachusetts Institute of Technology, publiziert, für deren Weiterentwicklung K. B. Mullis und M. Smith 1993 mit dem Nobelpreis für Chemie ausgezeichnet wurden. Seitdem wurde die Methode immer weiter optimiert und durch diverse Modifikationen erweitert. So können heute durch unterschiedliche PCR-Techniken qualitative, quantitative oder auch Fragestellungen der Genexpression bearbeitet werden.
(Hintergrundbild: © David Mencin/UNAVCO. Alle Rechte vorbehalten.)

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