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211.
Miguel F. Gonzales Teresa Brooks Stefan U. Pukatzki Daniele Provenzano 《Journal of visualized experiments : JoVE》2013,(80)
Electroporation has become a widely used method for rapidly and efficiently introducing foreign DNA into a wide range of cells. Electrotransformation has become the method of choice for introducing DNA into prokaryotes that are not naturally competent. Electroporation is a rapid, efficient, and streamlined transformation method that, in addition to purified DNA and competent bacteria, requires commercially available gene pulse controller and cuvettes. In contrast to the pulsing step, preparation of electrocompetent cells is time consuming and labor intensive involving repeated rounds of centrifugation and washes in decreasing volumes of sterile, cold water, or non-ionic buffers of large volumes of cultures grown to mid-logarithmic phase of growth. Time and effort can be saved by purchasing electrocompetent cells from commercial sources, but the selection is limited to commonly employed E. coli laboratory strains. We are hereby disseminating a rapid and efficient method for preparing electrocompetent E. coli, which has been in use by bacteriology laboratories for some time, can be adapted to V. cholerae and other prokaryotes. While we cannot ascertain whom to credit for developing the original technique, we are hereby making it available to the scientific community. 相似文献
212.
Sakine Simsekyilmaz Fabian Schreiber Stefan Weinandy Felix Gremse Tolga Taha S?nmez Elisa A. Liehn 《Journal of visualized experiments : JoVE》2013,(75)
Despite the considerable progress made in the stent development in the last decades, cardiovascular diseases remain the main cause of death in western countries. Beside the benefits offered by the development of different drug-eluting stents, the coronary revascularization bears also the life-threatening risks of in-stent thrombosis and restenosis. Research on new therapeutic strategies is impaired by the lack of appropriate methods to study stent implantation and restenosis processes. Here, we describe a rapid and accessible procedure of stent implantation in mouse carotid artery, which offers the possibility to study in a convenient way the molecular mechanisms of vessel remodeling and the effects of different drug coatings. 相似文献
213.
Ad Vos Tobias Nolden Christiane Habla Stefan Finke Conrad M. Freuling Jens Teifke Thomas Müller 《European Journal of Wildlife Research》2013,59(5):637-643
Raccoons can be found almost everywhere in Germany since their first successful introduction in 1934. Although the animal is a well-known reservoir species for rabies in the USA, during the last European fox rabies epizootic, only a few rabid raccoons were reported from Germany. In recent years, the raccoon population density has increased tremendously, especially in (semi) urban settings. Presently, Germany is free of terrestrial wildlife rabies. To assess the potential risk that the raccoon population in Germany could act as a reservoir species upon reemergence of rabies, the susceptibility of the local raccoon population was investigated. Wild-caught animals were inoculated with the most likely lyssavirus variants to infect the local population. It was shown that the raccoons were fully susceptible for a dog and raccoon rabies virus isolate. Also, five of six raccoons inoculated with a fox rabies virus isolate showed clinical signs. However, none of the raccoons infected with European Bat Lyssavirus type 1 succumbed to rabies; meanwhile, all these raccoons seroconverted. It is concluded that the highest risk for the raccoon population in Germany to become infected with lyssaviruses is through the importation of rabies infected dogs. 相似文献
214.
Marcus Bär Joachim Klaer Lothar Weinhardt Regan G. Wilks Stefan Krause Monika Blum Wanli Yang Clemens Heske Hans‐Werner Schock 《Liver Transplantation》2013,3(6):777-781
The surface properties of CuInS2 (CIS) thin‐film solar cell absorbers are investigated by a combination of electron and soft X‐ray spectroscopies. Spatially separated regions of varying colors are observed and identified to be dominated by either CuS or Cu2S surface phases. After their removal by KCN etching, the samples cannot be distinguished by eye and the CIS surface is found to be Cu‐deficient in both regions. However, a significantly more pronounced off‐stoichiometry in the region initially covered by Cu2S can be identified. In this region, the resulting surface band gap is also significantly larger than the EgSurf of the initially CuS‐terminated region. Such variations may represent a hidden parameter which, if overlooked, induces irreproducibility and thus prevents systematic optimization efforts. 相似文献
215.
Stefan Hindersin Marco Leupold Martin Kerner Dieter Hanelt 《Bioprocess and biosystems engineering》2013,36(3):345-355
Photosynthetic activity and temperature regulation of microalgal cultures (Chlorella vulgaris and Scenedesmus obliquus) under different irradiances controlled by a solar tracker and different cell densities were studied in outdoor flat panel photobioreactors. An automated process control unit regulated light and temperature as well as pH value and nutrient concentration in the culture medium. CO2 was supplied using flue gas from an attached combined block heat and power station. Photosynthetic activity was determined by pulse amplitude modulation fluorometry. Compared to the horizontal irradiance of 55 mol photons m?2 d?1 on a clear day, the solar tracked photobioreactors enabled a decrease and increase in the overall light absorption from 19 mol photons m?2 d?1 (by rotation out of direct irradiance) to 79 mol photons m?2 d?1 (following the position of the sun). At biomass concentrations below 1.1 g cell dry weight (CDW) L?1, photoinhibition of about 35 % occurred at irradiances of ≥1,000 μmol photons m?2 s?1 photosynthetic active radiation (PAR). Using solar tracked photobioreactors, photoinhibition can be reduced and at optimum biomass concentration (≥2.3 g CDW L?1), the culture was irradiated up to 2,000 μmol photons m?2 s?1 to overcome light limitation with biomass yields of 0.7 g CDW mol photons?1 and high photosynthetic activities indicated by an effective quantum yield of 0.68 and a maximum quantum yield of 0.80 (F v/F m). Overheating due to high irradiance was avoided by turning the PBR out of the sun or using a cooling system, which maintained the temperature close to the species-specific temperature optima. 相似文献
216.
217.
Riho Gross Stefan Palm Kuldar Kõiv Tore Prestegaard Japo Jussila Tiit Paaver Juergen Geist Harri Kokko Anna Karjalainen Lennart Edsman 《Conservation Genetics》2013,14(4):809-821
Noble crayfish (Astacus astacus L.), the most highly valued freshwater crayfish in Europe, is threatened due to a long-term population decline caused mainly by the spread of crayfish plague. Reintroduction of the noble crayfish into restored waters is a common practice but the geographic and genetic origin of stocking material has rarely been considered, partially because previous genetic studies have been hampered by lack of nuclear gene markers with known inheritance. This study represents the first large scale population genetic survey of the noble crayfish (633 adults from 18 locations) based on 10 newly developed microsatellite markers. We focused primarily on the Baltic Sea area (Estonia, Finland and Sweden) where the largest proportion of the remaining populations exists. To allow comparisons, samples from the Black Sea catchment (the Danube drainage) were also included. Two highly differentiated population groups were identified corresponding to the Baltic Sea and the Black Sea catchments, respectively. The Baltic Sea catchment populations had significantly lower genetic variation and private allele numbers than the Black Sea catchment populations. Within the Baltic Sea area, a clear genetic structure was revealed with population samples corresponding well to their geographic origin, suggesting little impact of long-distance translocations. The clear genetic structure strongly suggests that the choice of stocking material for re-introductions and supplemental releases needs to be based on empirical genetic knowledge. 相似文献
218.
Nicotiana attenuata HSPRO (NaHSPRO) is a negative regulator of seedling growth promoted by the fungus Piriformospora indica. Homologs of NaHSPRO in Arabidopsis thaliana (i.e., AtHSPRO1 and AtHSPRO2) are known to physically interact with the AKINβγ subunit of the SnRK1 complex.2 To investigate whether NaHSPRO is associated with SnRK1 function during the stimulation of seedling growth by P. indica, we studied N. attenuata plants silenced in the expression of NaGAL83 (as-gal83 plants)—a gene that encodes for the regulatory β-subunit of SnRK1—and plants silenced in the expression of both NaHSPRO and NaGAL83 (ir-hspro/as-gal83 plants). The results showed that P. indica differentially stimulated the growth of both as-gal83 and ir-hspro/as-gal83 seedlings compared with control seedlings, with a magnitude similar to that observed in ir-hspro seedlings. Thus, we showed that, similar to NaHSPRO, NaGAL83 is a negative regulator of seedling growth stimulated by P. indica. We propose that the effect of NaHSPRO on seedling growth is associated with SnRK1 signaling. 相似文献
219.
220.
Marc Baguelin Stefan Flasche Anton Camacho Nikolaos Demiris Elizabeth Miller W. John Edmunds 《PLoS medicine》2013,10(10)