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31.
When nucleosome cores reconstituted from chicken erythrocyte histones and a 160 bp DNA molecule are exposed to echinomycin, a bis-intercalating antitumour antibiotic, the DNA appears to rotate with respect to the histone octamer by about half a turn. New bands appear in patterns of DNAase I digestion at positions approximately mid-way between those characteristic of control core samples, while the control pattern is largely suppressed. Similar (but not identical) changes are produced when nucleosome cores are exposed to distamycin, a non-intercalating DNA-binding antibiotic. The effects of both ligands can be explained in terms of a change in rotational orientation of the core DNA, so as to place antibiotic binding sites on the inward-facing (concave) surface of the DNA supercoil. Presumably this serves to optimise non-bonded contacts with the polynucleotide backbone. These results establish that the positioning of DNA about the histone octamer is not absolutely determined by its nucleotide sequence, but may be modified by the binding of such relatively small molecules as antibiotics. 相似文献
32.
33.
Effect of metabolic inhibitors and temperature on uptake and translocation of ca and k by intact bean plants 总被引:1,自引:0,他引:1
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The dependence of Ca uptake and translocation by intact roots of Phaseolus vulgaris on concurrent root metabolism was investigated using 45Ca-labeled Hoagland solutions at one-half and one-twentieth strength (2.5 and 0.25 mM Ca2+). Adsorbed and absorbed 45Ca fractions in the roots were distinguished on the basis of the time course of exchange with the outer solution. Uptake of 42K, of which the characteristics are better known, was measured for comparison. The absorbed 45Ca fraction showed a markedly nonlinear increase with time in contrast to the near linear increase in 42K. Exposure of roots to cyanide, arsenate, 2,4-dinitrophenol, or low temperatures caused only slight reductions in 45Ca absorption by roots, but significant reductions of 42K. In all treatments involving inhibitors and low temperatures, the translocation to shoots of both 45Ca and 42K was strongly inhibited. The conclusion that much of the absorbed 45Ca fraction in the root tissue is taken up by processes which are not rate-limited by metabolism is discussed. 相似文献
34.
35.
ß-Glucuronidase (GUS) and chloramphenicol acetyl transferase (CAT) were used as reporter proteins in protoplasts from embryogenic suspension cultures of Picea glauca (Moench) Voss (white spruce). Plasmid DNA enclosing chimeric GUS and CAT constructs, using the cauliflower mosaic virus 35S promoter, was introduced into Picea glauca protoplasts using polyethylene glycol (PEG). Transient expression was detected 12 to 40 h after PEG-mediated DNA delivery. Dose-response curves using covalently closed circular plasmid DNA, in the absence of carrier DNA, have been obtained for each of these reporter genes. Linearized plasmid DNA gave lower levels of expression than covalently closed circular plasmid DNA when assayed 40 h after PEG-mediated DNA transfer. The use of carrier DNA (herring sperm DNA), in combination with covalently closed circular plasmid DNA, increased the level of expression of GUS by about 50%. CAT expression was enhanced if PEG-mediated delivery was performed on ice rather than at room temperature. The highest level of expression for CAT, and the lowest signal-to-noise ratio, was found 24 h after PEG-mediated DNA transfer. Both GUS and CAT provided results that were quantifiable and can therefore be used as reporter genes in Picea glauca.Abbreviations CAT
chloramphenicol acetyl transferase
- GUS
ß-glucuronidase
- CaMV
cauliflower mosaic virus
- NOS
nopaline synthase
- CCC
covalently closed circular DNA
- L
linear DNA
- PEG
polyethylene glycol
- HS
herring sperm DNA
- P
protoplasts
- PCM
protoplast culture medium
- MES
morpholinoethane-sulfonic acid
- Cm
chloramphenicol
- Ac
acetylated
- MUG
4-methyl umbelliferyl ß-D-glucuronide
- TLC
thin layer chromatography 相似文献
36.
37.
Characterization of gamma-aminobutyric acid and dopamine overflow following acute implantation of a microdialysis probe 总被引:7,自引:0,他引:7
The present study characterized the voltage and calcium dependence of gamma-aminobutyric acid and dopamine overflow after the acute implantation of a microdialysis probe. Probes were implanted in dorsolateral striatum and globus pallidus. Experiments were performed under light halothane anesthesia. Basal, extracellular levels of GABA were not affected by tetrodotoxin (TTX) and were increased to 140 percent of basal values by calcium free Ringer. Basal, extracellular levels of dopamine were reduced to 14 percent of basal values by the addition of TTX and to 30 percent of basal values by the removal of calcium from the Ringer solution. The results suggest that in this in vivo preparation basal extracellular dopamine is largely of vesicular origin while GABA is not. 相似文献
38.
J Drew A Liodakis R Chan H Du M Sadek R Brownlee W H Sawyer 《Biochemistry international》1990,22(6):983-992
Lipid emulsions consisting of a surface monolayer of phospholipid enclosing a core of neutral lipids have been prepared by repeated extrusion through polycarbonate filters of defined pore size. Particle size, as measured by photon correlation spectroscopy, decreases on successive passes through a 100 nm filter, reaching a near constant value (130-150 nm) after 4 passes. A corresponding decrease in the standard deviation of the particle size distribution occurs during this process. The recovery of lipids, especially of cholesterol and cholesterol ester, is improved if the emulsion is sonicated before extrusion through filters. [31P]-NMR and fluorescence techniques are used to confirm that the resulting structures are emulsions rather than lipid bilayers. 相似文献
39.
Richard W. Joyiv Trevor A. Thorpe 《In vitro cellular & developmental biology. Plant》1990,26(6):643-646
Summary The effects of excision, light and cytokinin (N6-benzyladenine) on14C-acetate metabolism in cotyledons ofPinus radiata (D. Don) were determined.14CO2 was released and the distribution of radioactivity into lipids, sugars, organic acids and amino acids was determined. While
light and cytokinin generally caused some increase in metabolism, the effect of excision, i.e., wounding, was most pronounced.
Specific metabolites examined (citrate, malate, succinate, alanine, aspartate, glutamate and glutamine) were at least 50%
greater in14C-labeling in excised cotyledons as compared to intact seedlings. This enhancement of wound metabolism would mask possible
morphogenically-related changes occurring at that time.
This research was supported by the Natural Sciences and Engineering Research Council of Canada Grant A-6467 to T.A. Thorpe. 相似文献
40.
Graham W. Burton Keith U. Ingold Kevin H. Cheeseman Trevor F. Slater 《Free radical research》1990,11(1):99-107
-Tocopherol, a superior chain-breaking, peroxyl radical-trapping antioxidant and the most active component of vitamin E, is elevated in liver tumor cells, contributing to their greater resistance towards lipid peroxidation compared to cells from normal tissues. Also, in regenerating rat liver the level of vitamin E has been found to fluctuate in phase with the rate of cell division. In order to study the biokinetcis and mechanisms of the distribution of vitamin E in organs and within tissues of animals, deuterated forms of -tocopherol have been synthesized and their uptake into blood and tissues has been measured by gas chromatography-mass spectrometry. Measurement of the competitive uptake from a mixture of the RRR-and SRR--tocopherol stereoisomers labelled with different amounts of deuterium shows that the liver exerts a strong preference for secretion of the natural (RRR) stereoisomer into the plasma. It is suggested that a tocopherol-binding protein plays a key role in this process. 相似文献