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1.
Herv Canut Johann Bauer Gerhard Weber 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》1999,722(1-2)
The review focuses on the multiple separating regimes that offers the free flow electrophoresis technique: free flow zone electrophoresis, isoelectric focusing, isotachophoresis, free flow step electrophoresis. Also, the feasibility to apply either interval or continuous flow electrophoresis is evaluated. The free flow zone electrophoresis regime is generally selected for the separation of cells, organelles and membranes while the other regimes find their largest fields of applications in the purification of proteins and peptides. The latter regimes present the highest resolution efficiency. Therefore, a large part of this review is devoted to the applicabilities of these different regimes to the purification of organelles and membrane vesicles at the preparative scale. Recent developments, both in instrumentation and procedures, are described. The major achievements in plant membrane fractionation obtained with free flow electrophoresis are outlined. The related procedures are both analytical and preparative: they separate tonoplast and plasma membrane simultaneously from the same homogenate, they discriminate for one type of membrane vesicles of opposite orientation, and process large quantities of membrane material by reason of the continuous flow mode. Recent advances using electromigration techniques that permit confirmation of the dynamic state of membranes, characterisation of complex membrane-dependent functions and discovery of new membrane-localised activities are presented. 相似文献
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The nucleotide sequence of a Sau3A1 restriction nuclease fragment that complemented an Escherichia coli chlD::Mu cts mutant strain was determined. DNA and deduced amino acid sequence analysis revealed two open reading frames (ORFs) that potentially codes for proteins with amino acid sequence homology with binding protein-dependent transport systems. One of the ORFs showed a sequence that encoded a protein with properties that were characteristic of a hydrophobic inner membrane protein. The other ORF, which was responsible for complementing a chlD mutant, encoded a protein with conserved sequences in nucleotide-binding proteins and hydrophilic inner membrane proteins in active transport systems. A proposal that the chlD locus is the molybdate transport operon is discussed in terms of the chlD phenotype. 相似文献
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Substrate specifity of the proton-driven hexose cotransport carrier in the plasmalemma of photoautotrophic suspension cells of Chenopodium rubrum L. has been studies through the short-term perturbation of 14C-labelled efflux of 3-O-methyl-d-glucose. Efflux, occurring exclusively via carrier-mediated exchange diffusion, is trans-stimulated by the substrate and trans-inhibited by the glucose-transport inhibitors phlorizin (K 1/2=7.9 mM) and its aglucon phloretin (K 1/2=84 μM); with both inhibitors, 3-O-methyl-d-glucose efflux may be blocked completely. Trans-stimulation of efflux (up to fourfold) by a variety of the d-enantiomers of neutral hexoses, including glucose (K 1/2=48 μM), 3-O-methyl-d-glucose (K 1/2=139 μM), and fructose (K 1/2=730 μM), but not by, for instance, d-allose, and l-sorbose, shows that carrier-substrate interaction critically involves the axial position at C-1 and C-3, respectively. We suggest that substrate binding by the Chenopodium hexose carrier involves both hydrophobic interaction with the pyran-ring and hydrogen-ion bonding at C-1 and C-3 of the d-glucose conformation. 相似文献
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Calli of P. argentatum were grown on a newly designed liquid nutrient flow-through system which facilitated the subculturing of calli and delayed
browning for 6 weeks. Friable calli were obtained on half-strength Gamborg B5-medium supplemented with 0.05 mgl−1 2,4-dichlorophenoxyacetic acid. Shoots developed on media supplemented with 0.2 mgl−1 benzylaminopurine but lacking 2,4-dichlorophenocyacetic acid. 相似文献
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The method of tight-seal whole-cell recording was used to study the amino-acid specificity of the Na+/alanine cotransporter in pancreatic acinar cells. Single cells or small clusters of electrically coupled cells were obtained by enzymatic dissociation of mouse pancreas. Inward currents were measured under 'zero-trans' conditions, i.e., at finite concentrations of Na+ and amino acid at the extracellular side and vanishing concentrations at the cytoplasmic side. The cotransporter, which corresponds to 'system A', as previously defined in the literature, was found to exhibit a wide tolerance to neutral amino acids (L-cysteine, L-serine, L-alanine, glycine, L-phenylalanine). Competition experiments with 2-methylaminoisobutyric acid (MeAIB) indicate that for glycine a second electrogenic transport system exists in pancreatic acinar cells. 相似文献
7.
Chromosomal in situ suppression hybridization of human gonosomes and autosomes and its use in clinical cytogenetics 总被引:5,自引:0,他引:5
A. Jauch C. Daumer P. Lichter J. Murken T. Schroeder-Kurth T. Cremer 《Human genetics》1990,85(2):145-150
Summary DNA libraries from sorted human gonosomes were used selectively to stain the X and Y chromosomes in normal and aberrant cultured human cells by chromosomal in situ suppression (CISS-) hybridization. The entire X chromosome was stained in metaphase spreads. Interphase chromosome domains of both the active and inactive X were clearly delineated. CISS-hybridization of the Y chromosome resulted in the specific decoration of the euchromatic part (Ypter-q11), whereas the heterochromatic part (Yq12) remained unlabeled. The stained part of the Y chromosome formed a compact domain in interphase nuclei. This approach was applied to amniotic fluid cells containing a ring chromosome of unknown origin (47,XY; +r). The ring chromosome was not stained by library probes from the gonosomes, thereby suggesting its autosomal origin. The sensitivity of CISS-hybridization was demonstrated by the detection of small translocations and fragments in human lymphocyte metaphase spreads after irradiation with 60Co-gamma-rays. Lymphocyte cultures from two XX-males were investigated by CISS-hybridization with Y-library probes. In both cases, metaphase spreads demonstrated a translocation of Yp-material to the short arm of an X chromosome. The translocated Y-material could also be demonstrated directly in interphase nuclei. CISS-hybridization of autosomes 7 and 13 was used for prenatal diagnosis in a case with a known balanced translocation t(7;13) in the father. The same translocation was observed in amniotic fluid cells from the fetus. Specific staining of the chromosomes involved in such translocations will be particularly important, in the future, in cases that cannot be solved reliably by conventional chromosome banding alone.Dedicated to Professor Friedrich Vogel on the occasion of his 65th birthday 相似文献
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Zusammenfassung Die Zellkerne vonPseudolysimachion spicatum, Veronica cymbalaria undV. gentianoides enthalten zweierlei Eiweißkörper von je nach dem Entwicklungszustand unterschiedlicher Konsistenz. Bis zum Höhepunkt ihrer Entwicklung liegen sie als anscheinend amorphe flüssige oder gelartige Gebilde vor. Später tritt — mit gewebespezifischen Unterschieden — Verfestigung und Kristallisation ein, und zwar bei beiden nicht gleichzeitig und z. T. auch nur beim großen. Beiderlei Körper unterscheiden sich deutlich in physikalischer und chemischer Hinsicht, außerdem in ihrer Entwicklungsgeschichte und Strukturveränderung. Sie fehlen in den Schließzellen, im Antherentapetum, in den Pollenmutterzellen, Pollenkörnern, im Embryosack, im Endosperm und im Embryo.Die Zellkerne in der Blattepidermis vonPenstemon barbatus enthalten ebenfalls zweierlei Eiweißkörper. Davon liegt einer als kugeliges Gebilde, der andere als Kristallstapel vor.
Summary In the nuclei ofPseudolysimachion spicatum, Veronica cymbalaria, V. gentianoides there occur two different kinds of protein bodies. They clearly differ from one another in physical and chemical regard as well as in their ontogeny. At first they are amorphous, liquid or geluous. As a rule a consolidation or crystallization with differences according to the tissue takes place after the climax of the development, and that not at the same time within both bodies. In the stomata cells, pollen mother cells, in the embryo sac, in the endosperm, in the endothelium and in the embryo both bodies do not occur.The nuclei in the epidermis of the leaves ofPenstemon barbatus also contain two different kinds of protein bodies, one of them appearing as a round body, and the other one as a pile of crystal plates.相似文献
10.
Age-specific life tables of two important pests of cowpea, Vigna unguiculata (L.) Walp., the pod sucking bugs Clavigralla tomentosicollis Stål and C. shadabi Dolling (Heteroptera: Coreidae), were obtained from observations carried out at different temperatures. A biophysical model was found satisfactory to describe the temperature-response of developmental and mortality rates of egg and nymphal stages, with a peak developmental rate around 34°C in both species. The variability in development times was small and the experimental data did not permit any conclusion with regard to the Erlang probability density function. Survival of eggs and nymphs remained high between 20° and 30°C for both species. At temperatures above 34°C, C. tomentosicollis survivorship and fecundity was higher than that of C. shadabi, which in turn laid more eggs at temperatures between 20° and 30°C. Maximum fecundity is estimated to be at 29°C for C. tomentosicollis (99 eggs/female) and 26°C for C. shadabi (261 eggs/female). At 30°C, the intrinsic rate of increase reached a maximum in both species, 0.152 per day for C. tomentosicollis and 0.145 per day for C. shadabi, and remained high for C. tomentosicollis until 36°C. C. tomentosicollis performed significantly better on pigeonpea, Cajanus cajan Millsp., than on cowpea at higher temperatures. 相似文献