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1.
Maximilian Schmid Bianca Dufner Julius Dürk Konstanze Bedal Kristina Stricker Lukas Ali Prokoph Christoph Koch Anja K. Wege Henner Zirpel Ger van Zandbergen Rupert Ecker Bogdan Boghiu Uwe Ritter 《PloS one》2015,10(10)
Characterization of host-pathogen interactions is a fundamental approach in microbiological and immunological oriented disciplines. It is commonly accepted that host cells start to change their phenotype after engulfing pathogens. Techniques such as real time PCR or ELISA were used to characterize the genes encoding proteins that are associated either with pathogen elimination or immune escape mechanisms. Most of such studies were performed in vitro using primary host cells or cell lines. Consequently, the data generated with such approaches reflect the global RNA expression or protein amount recovered from all cells in culture. This is justified when all host cells harbor an equal amount of pathogens under experimental conditions. However, the uptake of pathogens by phagocytic cells is not synchronized. Consequently, there are host cells incorporating different amounts of pathogens that might result in distinct pathogen-induced protein biosynthesis. Therefore, we established a technique able to detect and quantify the number of pathogens in the corresponding host cells using immunofluorescence-based high throughput analysis. Paired with multicolor staining of molecules of interest it is now possible to analyze the infection profile of host cell populations and the corresponding phenotype of the host cells as a result of parasite load. 相似文献
2.
The temperate phage P1 encodes two genes whose products antagonize the action of the phage's C1 repressor of lytic functions, namely a distantly linked antirepressor gene, ant, and a closely linked c1 inactivator gene, coi. Starting with an inducible coi-recombinant plasmid, Coi protein was overproduced and purified to near homogeneity. By using a DNA mobility shift assay we demonstrate that Coi protein inhibits the operator binding of the C1 repressors of the closely related P1 and P7 phages. Coi protein (Mr = 7,600) exerts its C1-inactivating function by forming a complex with the C1 repressor (Mr = 32,500) at a molar ratio of about 1:1, as shown by density gradient centrifugation and gel filtration. C1 repressor and Coi protein are recovered in active form from the complex, suggesting that noncovalent interactions are the sole requirements for complex formation. The interplay of repressor and antagonists operating in the life cycle of P1 is discussed. 相似文献
3.
R Chakrabarti N E Pfeiffer D E Wylie S M Schuster 《The Journal of biological chemistry》1989,264(14):8214-8221
Incorporation of asparagine synthetase-specific monoclonal antibodies into L5178Y D10/R (L-asparaginase-resistant) murine lymphoma cells by osmotic lysis of pinocytic vesicles was used to evaluate the potential of the technique for macromolecular incorporation for metabolic studies. Nonspecific effects of the incorporation procedure included temporary inhibition of protein and DNA synthesis by 80-85% and a transitory loss of membrane integrity. Cells incorporated with an antibody inhibitory to tumor cell asparagine synthetase showed increased dependence upon an exogenous source of asparagine in the culture medium, while cells incorporated with a control antibody were not affected. These studies demonstrated that incorporation of inhibitory monoclonal antibodies into cells can be used to study the short term metabolic role of specific enzymes; however, the metabolic effects induced by the specific macromolecule must be evaluated within the context of the nonspecific effects caused by the osmotic treatment required for incorporation. 相似文献
4.
The open reading frame encoding ATPase subunit 6 in Oenothera mitochondria is edited at 21 positions in all cDNA clones investigated. Only one of these events is silent, all others improve similarity between the homologous polypeptides of other species. The introduction of a new UAA termination codon shortens the polypeptide by 35 amino acids to a carboxy terminus conserved in other species. In one of the cDNA clones, an additional editing event was observed resulting in a premature UAA termination codon in the amino terminal region. 相似文献
5.
The flowering time is the most susceptible period for primary infection of wheat heads byFusarium spp. During this period spores can be deposited into the opened wheat florets where they may later cause infections. We quantitatively
explored the relationship between variables related to the flowering process and the infection level byFusarium graminearum in single spikelets. We imitated open (chasmogamous) and closed (cleistogamous) flowering by injecting well-defined amounts
of spores into and between wheat florets. Applying the spores between the florets resulted in weaker disease symptoms and
significantly lower amounts ofFusarium mycotoxins. With larger numbers of spores, the disease symptoms became more pronounced and the mycotoxin amounts per spikelet
increased significantly.
Our results indicate that the probability of primary infection is approximately proportional to the number of spores reaching
the open florets during the flowering process. The breeding of wheat lines which flower partially or completely cleistogamously
might reduce theFusarium susceptibility in wheat. 相似文献
6.
7.
Schuster Martina Tewary Gargi Bao Xuanwen Subedi Prabal Hauck Stefanie M. Olsen Ann Karin Eide Dag Markus Trott Klaus Rüdiger Götz Sebastian Atkinson Michael J. Rosemann Michael 《Radiation and environmental biophysics》2021,60(4):689-689
Radiation and Environmental Biophysics - 相似文献
8.
Feed grain production for on-farm use is widespread in Bavarian agriculture and garants a reliable production of healthy foodstuff. Therefore the quality of feed grain is very important for farmers. In an orientating investigation of the on-farm stored feed grain quality from the harvest years 1991 to 2000 1757 samples were analysed. Based on values for guidance by the German government for DON and ZEA and Commission Regulation EC No 472/2002 for OTA the results show that only 2% of the positive samples have amounts higher then 0,05 mg ZEA/kg grain, 4% of the positive samples contain more then 1 mg DON/kg grain and 2% of the positive samples have amounts higher then 0,003 mg OTA/kg feed grain. These illustrate the good conservation and storage quality on Bavarian farms. 相似文献
9.
Hartmut Heller Sebastian Brandt Klaus-Dieter Schuster 《Journal of applied physiology》2002,92(4):1770; author reply 1770-1770; author reply 1771
10.