SummaryMiraculin is a taste-modifying protein isolated from the red berries of Richadella dulcifica , a shrub native to West Africa. Miraculin by itself is not sweet, but it is able to turn a sour taste into a sweet taste. This unique property has led to increasing interest in this protein. In this article, we report the high-yield production of miraculin in transgenic tomato plants. High and genetically stable expression of miraculin was confirmed by Western blot analysis and enzyme-linked immunosorbent assay. Recombinant miraculin accumulated to high levels in leaves and fruits, up to 102.5 and 90.7 µ g/g fresh weight, respectively. Purified recombinant miraculin expressed in transgenic tomato plants showed strong sweetness-inducing activity, similar to that of native miraculin. These results demonstrate that recombinant miraculin was correctly processed in transgenic tomato plants, and that this production system could be a good alternative to production from the native plant.
As metabolomics can provide a biochemical snapshot of an organism's phenotype it is a promising approach for charting the unintended effects of genetic modification. A critical obstacle for this application is the inherently limited metabolomic coverage of any single analytical platform. We propose using multiple analytical platforms for the direct acquisition of an interpretable data set of estimable chemical diversity. As an example, we report an application of our multi-platform approach that assesses the substantial equivalence of tomatoes over-expressing the taste-modifying protein miraculin. In combination, the chosen platforms detected compounds that represent 86% of the estimated chemical diversity of the metabolites listed in the LycoCyc database. Following a proof-of-safety approach, we show that % had an acceptable range of variation while simultaneously indicating a reproducible transformation-related metabolic signature. We conclude that multi-platform metabolomics is an approach that is both sensitive and robust and that it constitutes a good starting point for characterizing genetically modified organisms.
Variability of R-R intervals and arterial blood pressure signals in chronically instrumented fetal lambs was analyzed by power spectral analysis based on an assumption of maximum entropy. There were four consistent components, very low (VL, 0.01-0.025 cycle/beat), low (L, 0.025-0.125 cycle/beat), middle (M, 0.125-0.2 cycle/beat), and high (H, 0.2-0.5 cycle/beat), in the normal heart rate variability and blood pressure spectra. Integrated peaks in the power spectrum were compared before and after the administration of sympathetic and parasympathetic blockades. beta-Sympathetic blockade reduced the spectral power in the VL and L frequency components. alpha-Sympathetic blockade reduced only the M frequency component in the spectrum of R-R interval variability. Parasympathetic blockade reduced the H and L frequency components in the R-R interval variability spectrum but increased these components in the systolic blood pressure variability spectrum. The results clearly demonstrate the association between fetal autonomic activity and change of power spectrum of heart rate and blood pressure variability.
Well-defined
monolayers with single-crystalline-like molecular
arrangements of dinaphtho[2,3-b:2′,3′-f]thieno[3,2-b]-thiophene (DNTT) and picene, which are a new class of
organic semiconductors with enhanced intermolecular interactions,
were fabricated and characterized. Although both molecules initially
form a loosely packed monolayer with a flat-lying molecule, they undergo
phase transition into a densely packed monolayer with single-crystalline-like
molecular arrangements with increasing molecular density. Upon the
phase transition of the monolayer, the highest occupied molecular
orbital (HOMO) level of these molecules splits into two features,
as suggested from both the ultraviolet photoelectron spectroscopy
and density functional theory calculations. The splitting of the HOMO
level was observed to be similar to that expected for the molecular
arrangement in the single crystal. This splitting, which has not been
observed in the polycrystalline film, suggests a substantial overlap
of the HOMO in the well-ordered monolayers.
Adsorptions of alkali metals (such as K and Li) on monolayers of coronene and picene realize the formation of ordered phases, which serve as well-defined model systems for metal-intercalated aromatic superconductors. Upon alkali-doping of the monolayers of coronene and picene, scanning tunneling microscopy and X-ray absorption spectroscopy revealed the rearrangement of the entire molecular layer. The K-induced reconstruction of both monolayers resulted in the formation of a structure with a herringbone-like arrangement of molecules, suggesting the intercalation of alkali metals between molecular planes. Upon reconstruction, a shift in both the vacuum level and core levels of coronene was observed as a result of a charge transfer from alkali metals to coronene. In addition, a new density of states near the Fermi level was formed in both the doped coronene and the doped picene monolayers. This characteristic electronic feature of the ordered monolayer has been also reported in the multilayer picene films, ensuring that the present monolayer can model the properties of the metal-intercalated aromatic hydrocarbons. It is suggested that the electronic structure near the Fermi level is sensitive to the molecular arrangement, and that both the strict control and determinations of the molecular structure in the doped phase should be important for the determination of the electronic structure of these materials.
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