Mannheim, Germany
Mannheim, Germany

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End C.,Promega GmbH | Walczuch C.,Promega GmbH | Buntru M.,Fraunhofer Institute for Molecular Biology and Applied Ecology
BioSpektrum | Year: 2014

Cell-free protein synthesis (CFPS) is an important tool for molecular biology and can be used in applied and basic research. Especially as a high throughput technology for functional genomics and proteomics, CFPS systems offer a significant advantage compared to living cells. Toxic, membrane or viral proteins can be readily expressed in vitro. However, a low synthesis performance currently limits the use for bioproduction purposes. To overcome this limitation, new systems are being developed. © 2014 Springer-Verlag Berlin Heidelberg. Literatur:.

Moritz W.,InSphero | Hanke S.,Promega GmbH | Kofer J.,Promega GmbH
BioSpektrum | Year: 2012

In higher order organisms, cell-to-cell interactions are crucial to fulfill concerted, tissue- or organ-specific tasks. Three-dimensional cell culture models reflect the natural environment of tissue structures in a more realistic way than cells in traditional monolayer cultures. By combining an automatable microtissue technology with a cell based assay, cell viability, cytotoxicity and the number of cells can be determined in complex three-dimensional cell cultures. © Springer-Verlag 2012.

Willmann M.,University of Cologne | Gerlach N.,University of Cologne | Buer B.,University of Cologne | Polatajko A.,University of Cologne | And 8 more authors.
Frontiers in Plant Science | Year: 2013

Arbuscular mycorrhizal fungi (AMF) form a mutually beneficial symbiosis with plant roots providing predominantly phosphorus in the form of orthophosphate (Pi) in exchange for plant carbohydrates on low P soils. The goal of this work was to generate molecular-genetic evidence in support of a major impact of the mycorrhizal Pi uptake (MPU) pathway on the productivity of the major crop plant maize under field and controlled conditions. Here we show, that a loss-of-function mutation in the mycorrhiza-specific Pi transporter gene Pht1;6 correlates with a dramatic reduction of above-ground biomass and cob production in agro-ecosystems with low P soils. In parallel mutant pht1;6 plants exhibited an altered fingerprint of chemical elements in shoots dependent on soil P availability. In controlled environments mycorrhiza development was impaired in mutant plants when grown alone. The presence of neighboring mycorrhizal nurse plants enhanced the reduced mycorrhiza formation in pht1;6 roots. Uptake of 33P-labeled orthophosphate via the MPU pathway was strongly impaired in colonized mutant plants. Moreover, repression of the MPU pathway resulted in a redirection of Pi to neighboring plants. In line with previous results, our data highlight the relevance of the MPU pathway in Pi allocation within plant communities and in particular the role of Pht1;6 for the establishment of symbiotic Pi uptake and for maize productivity and nutritional value in low-input agricultural systems. In a first attempt to identify cellular pathways which are affected by Pht1;6 activity, gene expression profiling via RNA-Seq was performed and revealed a set of maize genes involved in cellular signaling which exhibited differential regulation in mycorrhizal pht1;6 and control plants. The RNA data provided support for the hypothesis that fungal supply of Pi and/or Pi transport across Pht1;6 affects cell wall biosynthesis and hormone metabolism in colonized root cells. © 2013 Willmann, Gerlach, Buer, Polatajko, Nagy, Koebke, Jansa, Flisch and Bucher.

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