TY  - JOUR
A1  - Klamp, Tobias
A1  - Camps, Marta
A1  - Nieto, Benjamin
A1  - Guasch, Francesc
A1  - Ranasinghe, Rohan T.
A1  - Wiedemann, Jens
A1  - Petrášek, Zdeněk
A1  - Schwille, Petra
A1  - Klenerman, David
A1  - Sauer, Markus
T1  - Highly Rapid Amplification-Free and Quantitative DNA Imaging Assay
JF  - Scientific Reports
N2  - There is an urgent need for rapid and highly sensitive detection of pathogen-derivedDNAin a point-of-care (POC) device for diagnostics in hospitals and clinics. This device needs to work in a ‘sample-in-result-out’ mode with minimum number of steps so that it can be completely integrated into a cheap and simple instrument. We have developed a method that directly detects unamplified DNA, and demonstrate its sensitivity on realistically sized 5 kbp targetDNA fragments of Micrococcus luteus in small sample volumes of 20 mL. The assay consists of capturing and accumulating of target DNA on magnetic beads with specific capture oligonucleotides, hybridization of complementary fluorescently labeled detection oligonucleotides, and fluorescence imaging on a miniaturized wide-field fluorescence microscope. Our simple method delivers results in less than 20 minutes with a limit of detection (LOD) of,5 pMand a linear detection range spanning three orders of magnitude.
KW  - laboratory techniques and procedures
KW  - diseases
KW  - infectious diseases
KW  - assay systems
Y1  - 2013
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-130500
VL  - 3
IS  - 1852
ER  - 
TY  - JOUR
A1  - Wolf, Annette
A1  - Akrap, Nina
A1  - Marg, Berenice
A1  - Galliardt, Helena
A1  - Heiligentag, Martyna
A1  - Humpert, Fabian
A1  - Sauer, Markus
A1  - Kaltschmidt, Barbara
A1  - Kaltschmidt, Christian
A1  - Seidel, Thorsten
T1  - Elements of Transcriptional Machinery Are Compatible among Plants and Mammals
JF  - PLoS ONE
N2  - In the present work, the objective has been to analyse the compatibility of plant and human transcriptional machinery. The experiments revealed that nuclear import and export are conserved among plants and mammals. Further it has been shown that transactivation of a human promoter occurs by human transcription factor NF-\(\kappa\) B in plant cells, demonstrating that the transcriptional machinery is highly conserved in both kingdoms. Functionality was also seen for regulatory elements of NF-\(\kappa\) B such as its inhibitor I\(\kappa\)B isoform \(\alpha\) that negatively regulated the transactivation activity of the p50/RelA heterodimer by interaction with NF-\(\kappa\)B in plant cells. Nuclear export of RelA could be demonstrated by FRAP-measurements so that RelA shows nucleo-cytoplasmic shuttling as reported for RelA in mammalian cells. The data reveals the high level of compatibility of human transcriptional elements with the plant transcriptional machinery. Thus, Arabidopsis thaliana mesophyll protoplasts might provide a new heterologous expression system for the investigation of the human NF-\(\kappa\)B signaling pathways. The system successfully enabled the controlled manipulation of NF-\(\kappa\)B activity. We suggest the plant protoplast system as a tool for reconstitution and analyses of mammalian pathways and for direct observation of responses to e. g. pharmaceuticals. The major advantage of the system is the absence of interference with endogenous factors that affect and crosstalk with the pathway.
KW  - complexes
KW  - in vivo
KW  - DNA-binding
KW  - nuclear proe
KW  - gene expression
KW  - NF-KAPPA-B
KW  - RNA-binding protein
KW  - alpha
KW  - inflammation
KW  - homodimers
Y1  - 2013
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-131203
VL  - 8
IS  - 1
ER  -