Einfluss variabler Translationsgeschwindigkeit auf die Zelle(S. 452)
Andreas Czech, Frauke Adamla, Zoya Ignatova
Translation of mRNA into an amino acid sequence is mediated by ribosomes. tRNAs deliver amino acids to the growing polypeptide chain by pairing to the cognate codons of the mRNA. The concentration of cellular tRNAs is not uniform and varies upon development, disease and environmental stress. The different availability of tRNAs affects the ribosomal speed and hence protein production.
Kohlenhydratstoffwechsel in Archaea – Besonderheiten und Herausforderung(S. 455)
Archaea exhibit unique features especially with respect to their central carbohydrate metabolism characterized by many unusual pathways and enzymes with specific energetic and regulatory properties. This probably contributes to “metabolic thermoadaptation” towards high temperature environments in which many archaea reside. The unusual metabolic and adaptive properties of Archaea also possess a great potential for biotechnology, which becomes more and more accessible by systems biology approaches.
Retrotranslokation über die ER-Membran durch die Ubiquitinligase Hrd1(S. 459)
Proteins that fail to fold correctly in the endoplasmic reticulum (ER) are retrotranslocated back into the cytosol where they become ubiquitinated and are eventually degraded by the proteasome. This process is called ER-associated protein degradation (ERAD). Recent research revealed that the ER-membrane protein Hrd1, an E3 ubiquitin ligase, forms a minimal system to translocate misfolded proteins across the membrane of the ER.
Organohalid-Atmung bei Mikroorganismen(S. 462)
Torsten Schubert, Lorenz Adrian, Gabriele Diekert
Reductive dehalogenation is coupled to energy conservation via electron transport phosphorylation in organohalide-respiring bacteria. The electron transfer to the terminal reductase, the corrinoid-containing reductive dehalogenase, leads to the formation of a proton gradient across the membrane. The architecture of the electron transfer chains differs between different phyla. Organohalide respiration is an important part of the global halogen cycle and of high relevance in bioremediation.
Protein-Nukleinsäure-Interaktionen im Massenspektrometer(S. 466)
Alexandra Stützer, Aleksandar Chernev, Henning Urlaub
Protein-nucleic acid complexes play a significant role in all DNA- and RNArelated cellular processes. The identification of protein-nucleic acid interaction sites is important to understand the structure, function and regulation of these complexes. Here, new approaches in mass spectrometry using UV-induced cross-linking of nucleic acids with proteins have recently proven as an important technique to not only identify proteins but specific regions of proteins that interact with RNA or DNA.
Analyse des Mikromilieus künstlicher Gewebe mit Protein-Multiplex-Assays(S. 471)
Stefanie Thoms, Rebecca Jonczyk, Antonina Lavrentieva, Cornelia Blume
The development of bioartificial tissues using stem cells has an enormous potential in medicine. To determine effects of differentiation strategies on cell micro-milieus, a fast and efficient method is necessary. The protein multiplex assay is based on enzyme-linked immunosorbent assays and allows automated, quantitative and qualitative analysis of multiple proteins in small sample volumes. Here, this method is shown to verify differentiation of mesenchymal stem cells into endothelial cells.
Automatische Inspektion von Kulturschalen mit der PetriJet-Plattform(S. 476)
Felix Lenk, Patrick Oberthür, Christoph Otto
Benchtop lab automation systems address specific, recurring tasks in small or mid-sized laboratories. With the versatile, standardized PetriJet platform technology the handling of culture dishes has been automated completely. Through a range of different processing stations different tasks such as multi-perspective imaging can be carried out without user interference and with a higher throughput. The integrated sorting function and the LIMS-connectivity allow to speed up subsequent processes.
(Teil-)Automatisierung der liquid-overlay-Sphäroidkultivierung(S. 480)
Alexander Krüger, Melanie Hüther, Mirjam Ingargiola, Leoni A. Kunz-Schughart
3D tumor spheroid models are increasingly applied in secondary, cellbased drug testing due to certain therapy-relevant in vivo-like characteristics. Automation of routine and delicate steps in spheroid culturing and processing, including the gentle well-to-well transfer of spheroids, facilitates spheroid-based assay standardization. Semi-automated workflows minimize experimental variations and are recommended in particular for combination treatment testing and long-term monitoring.
Anwendungen & Produkte
Automatisierung für die personalisierte Krebstherapie(S. 496)
Manal Hadenfeld, Georg Rauser, Ulrike Köhl, Hinrich Abken, Michael Apel
Every year, in Germany about 17.000 patients are diagnosed with malignant melanoma, and 3.000 patients die as a consequence of the disease. Recently, many successful results in cancer therapy were achieved by the CAR T cell therapy. In the BMBF-funded project CD20 CAR-TIME, CAR T cells will be applied to treat melanoma patients in a clinical study. Moreover, this project consortium will explore how automation can de - liver this personalized form of therapy to a large number of patients.
Aufklärung toxischer Wirkmechanismen mittels Hochdurchsatz-RT-qPCR(S. 499)
Andrea Hartwig, Bettina Maria Fischer
Gene expression studies have become valuable tools to identify the complex mode of action of toxic or carcinogenic substances. In our group we established a quantitative high-throughput RT-qPCR technique using Fluidigm dynamic arrays on the BioMark™ System to analyze the expression level of 95 genes in 96 samples in parallel. By creating a special gene set comprising critical genes of the DNA damage response system the impact on genomic stability can be investigated.
Plattformunabhängige Isolierung hochmolekularer Nukleinsäuren(S. 501)
Melanie Kelm, Matthias Ludwig
More than 35 years after describing the first silica-based DNA and RNA isolation technique, novel methods in the field of nucleic acid extraction are invented now. The new method is especially in keeping with the trend toward automating processes and fullfil the need for qualitative, high molecular weight DNA, which gain importance in Next Generation Sequencing (NGS).
Kombinatorische Chemie im hochdichten Arrayformat(S. 532)
Felix Löffler, Frank Breitling, Alexander Nesterov-Müller
We synthesize peptide arrays by using laser pulses to transfer amino acids onto a synthesis slide that are embedded in a polymer. After patterning these razor-thin spots, the coupling starts by heating the polymer that now serves as a solvent to couple monomers to the surface. The main advantages of this nanolayer solid phase chemistry are the high degree of automation, the very high spot density, and the in situ activation of monomers due to mixing different reagents in the stacked nanolayers.
Immer (nur) Bioplastik? Cupriavidus necator als Produktionsplattform(S. 535)
Jessica Eggers, Stephan Hetzler, Alexander Steinbüchel
The facultative chemolithoautotrophic bacterium Cupriavidus necator is known for its ability to store large amounts of polyhydroxyalkanoates (PHA). However, its versatile metabolism, non-pathogenicity, genetic accessibility and productivity have promoted research for numerous value added products beyond PHA. With recent developments in meta - bolic engineering, it has the potential as an alternative production strain for various high value products in biotechnology.
Karriere, Köpfe & Konzepte
Die Universität Leipzig ehrt Rudolf Schönheimer(S. 542)
Am 1. April 2016 wurde das Institut für Biochemie der Medizinischen Fakultät der Universität Leipzig (bis 1985 „Physiologisch-Chemisches Institut“) 100 Jahre alt. Aus diesem Anlass wird ihm im Rahmen einer Feierstunde am 19. September 2016 der Name „Rudolf-Schönheimer- Institut für Biochemie“ verliehen. Damit ehrt die Universität Leipzig einen großen Wissenschaftler, der in diesem Institut von 1924 bis 1926 eine vertiefte Ausbildung in Physiologischer Chemie erhielt, 1933 aber Deutschland infolge seiner jüdischen Abstammung verlassen musste und in den USA zu Weltruhm gelangte.
Zellfreie Proteinsynthese: veraltete Technik oder moderne Methode?(S. 546)
Daniel Kokotek, Anna Dreyer, Sabrina Fischer, Alexander Gräwe, Luisa Keilholz, Janina Lüders
Modern diagnostics are inconceivable without quick and robust tests. With today’s lab-on-a-chip applications, cell-free expression comes back into focus. Two German student teams from the Universities of Bielefeld and Freiburg developed new diagnostic tools based on cell-free expression. Freiburg established a label-free screening for diseases; Bielefeld focused on the detection of harmful substances in water. Both systems offer robust and user-friendly alternatives to current diagnostic tools.