60 research outputs found
Correlation between morphology and magnetic properties of electrochemically produced cobalt powder particles
Cobalt 3D powder particles were successfully prepared by galvanostatic electrodeposition. The electrodeposited cobalt powders were characterized by X-ray diffraction (XRD) analysis, scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS) and SQUID magnetometry. It was shown that the morphology, structure and magnetic properties of cobalt particles were closely associated and could be easily controlled by adjusting the electrodeposition process parameters. The morphology of cobalt powder particles was strongly affected by the hydrogen evolution reaction as a parallel reaction to cobalt electrodeposition. Depending on the applied current density, two types of powder particles were formed: dendrites at lower and spongy-like particles at higher current densities. Morphologies and structures of powder particles were correlated with their magnetic properties, and compared with those of the bulk cobalt. In comparison with the properties of bulk cobalt, the obtained 3D structutes exhibited a decreased saturation magnetization (M-S), but an enhanced coercivity (H-C), which was explained by their peculiar morphology
Characterizing RecA-Independent Induction of Shiga toxin2-Encoding Phages by EDTA Treatment
Background: The bacteriophage life cycle has an important role in Shiga toxin (Stx) expression. The induction of Shiga toxin-encoding phages (Stx phages) increases toxin production as a result of replication of the phage genome, and phage lysis of the host cell also provides a means of Stx toxin to exit the cell. Previous studies suggested that prophage induction might also occur in the absence of SOS response, independently of RecA. Methodology/Principal Findings: The influence of EDTA on RecA-independent Stx2 phage induction was assessed, in laboratory lysogens and in EHEC strains carrying Stx2 phages in their genome, by Real-Time PCR. RecA-independent mechanisms described for phage l induction (RcsA and DsrA) were not involved in Stx2 phage induction. In addition, mutations in the pathway for the stress response of the bacterial envelope to EDTA did not contribute to Stx2 phage induction. The effect of EDTA on Stx phage induction is due to its chelating properties, which was also confirmed by the use of citrate, another chelating agent. Our results indicate that EDTA affects Stx2 phage induction by disruption of the bacterial outer membrane due to chelation of Mg 2+. In all the conditions evaluated, the pH value had a decisive role in Stx2 phage induction. Conclusions/Significance: Chelating agents, such as EDTA and citrate, induce Stx phages, which raises concerns due to their frequent use in food and pharmaceutical products. This study contributes to our understanding of the phenomenon o
Seamless Insert-Plasmid Assembly at High Efficiency and Low Cost
Seamless cloning methods, such as sequence- and ligation-independent cloning (SLIC) or the Gibson assembly, are essential tools for the construction of protein expression plasmids. We here show that single-stranded gaps in double-stranded plasmids, which for example occur in typical SLIC protocols, can drastically decrease the efficiency at which the DNA transforms competent E. coli bacteria. Conversely, filling-in of single-stranded gaps using DNA polymerase resulted in increased transformation efficiency. Ligation of the remaining nicks did not lead to a further increase in transformation efficiency. These data point out a critical factor for robust seamless cloning. Highly efficient insert-plasmid assembly can be achieved by using only T5 exonuclease and Phusion DNA polymerase, without Taq DNA ligase from the original Gibson protocol, which significantly reduces the cost of the reactions. We successfully used this method with two short insert-plasmid overlap regions, each counting only 15 nucleotides
Combinatorial optimization of synthetic operons for the microbial production of p-coumaryl alcohol with Escherichia coli
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Interaction between plant species and substrate type in the removal of CO2 indoors
Elevated indoor concentrations of carbon dioxide [CO2] cause health issues, increase workplace absenteeism and reduce cognitive performance. Plants can be part of the solution, reducing indoor [CO2] and acting as a low-cost supplement to building ventilation systems.
Our earlier work on a selection of structurally and functionally different indoor plants identified a range of leaf-level CO2 removal rates, when plants were grown in one type of substrate. The work presented here brings the research much closer to real indoor environments by investigating CO2 removal at a whole-plant level and in different substrates. Specifically, we measured how the change of growing substrate affects plants’ capacity to reduce CO2 concentrations. Spathiphyllum wallisii 'Verdi', Dracaena fragrans 'Golden Coast' and Hedera helix, representing a range of leaf types and sizes and potted in two different substrates, were tested. Potted plants were studied in a 0.15 m3 chamber under ‘very high’ (22000 lux), ‘low’ (~ 500 lux) and ‘no’ light (0 lux) in ‘wet’ (> 30 %) and ‘dry’ (< 20 %) substrate.
At ‘no’ and ‘low’ indoor light, houseplants increased the CO2 concentration in both substrates; respiration rates, however, were deemed negligible in terms of the contribution to a room-level concentration, as they added ~ 0.6% of a human’s contribution. In ‘very high’ light D. fragrans, in substrate 2, showed potential to reduce [CO2] to a near-ambient (600 ppm) concentration in a shorter timeframe (12 hrs, e.g. overnight) and S. wallisii over a longer period (36 hrs, e.g. weekend)
Combinatorial optimization of synthetic operons for the microbial production of p-coumaryl alcohol with Escherichia coli
BACKGROUND: Microbes are extensively engineered to produce compounds of biotechnological or pharmaceutical interest. However, functional integration of synthetic pathways into the respective host cell metabolism and optimization of heterologous gene expression for achieving high product titers is still a challenging task. In this manuscript, we describe the optimization of a tetracistronic operon for the microbial production of the plant-derived phenylpropanoid p-coumaryl alcohol in Escherichia coli. RESULTS: Basis for the construction of a p-coumaryl alcohol producing strain was the development of Operon-PLICing as method for the rapid combinatorial assembly of synthetic operons. This method is based on the chemical cleavage reaction of phosphorothioate bonds in an iodine/ethanol solution to generate complementary, single-stranded overhangs and subsequent hybridization of multiple DNA-fragments. Furthermore, during the assembly of these DNA-fragments, Operon-PLICing offers the opportunity for balancing gene expression of all pathway genes on the level of translation for maximizing product titers by varying the spacing between the Shine-Dalgarno sequence and START codon. With Operon-PLICing, 81 different clones, each one carrying a different p-coumaryl alcohol operon, were individually constructed and screened for p-coumaryl alcohol formation within a few days. The absolute product titer of the best five variants ranged from 48 to 52 mg/L p-coumaryl alcohol without any further optimization of growth and production conditions. CONCLUSIONS: Operon-PLICing is sequence-independent and thus does not require any specific recognition or target sequences for enzymatic activities since all hybridization sites can be arbitrarily selected. In fact, after PCR-amplification, no endonucleases or ligases, frequently used in other methods, are needed. The modularity, simplicity and robustness of Operon-PLICing would be perfectly suited for an automation of cloning in the microtiter plate format. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12934-015-0274-9) contains supplementary material, which is available to authorized users
Dynamics of seed protein biosynthesis in two soybean genotypes differing in drought susceptibility
The dynamics of seed storage protein biosynthesis was studied under field conditions during two vegetative seasons. Two soybean (Glycine max L. Merr.) genotypes were examined: BOS-A (drought tolerant) and L 121 (drought susceptible). Seed samples were taken from plants at three stages of seed maturation (50 and 70 d after flowering, and at full maturity). The earlier synthesis of the beta-subunit of the 7S protein occurred in the drought susceptible cultivar. We have not found such differences in the synthesis of the alpha- and alpha'-subunits of the 7S protein. Our results did not confirm significant genotypic differences in protein composition of the mature seeds between the cultivars studied, but have pointed out to the differences in the dynamics of protein biosynthesis during seed maturation and desiccation
Simultaneous determination of phosphorus-32 and calcium-45 activity in biological samples
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