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21.
Molecular cloning and nucleotide sequences of cDNAs specific for rat liver ribosomal proteins S17 and L30 总被引:10,自引:0,他引:10
O Nakanishi M Oyanagi Y Kuwano T Tanaka T Nakayama H Mitsui Y Nabeshima K Ogata 《Gene》1985,35(3):289-296
cDNA clones coding for rat liver ribosomal proteins S17 and L30 have been isolated by positive hybridization-translation assay from a cDNA library prepared from 8-9S poly(A)+RNA from free polysomes of regenerating rat liver. The cDNA clone specific for S17 protein (pRS17-2) has a 466-bp insert with the poly(A) tail. The complete amino acid (aa) sequence of S17 protein was deduced from the nucleotide sequence of the cDNA. S17 protein consists of 134 aa residues with an Mr of 15 377. The N-terminal aa sequence of S17 protein determined by automatic Edman degradation is consistent with the sequence data. The aa sequence of S17 shows strong homology (76.9%) to that of yeast ribosomal protein 51 [Teem and Rosbash, Proc. Natl. Acad. Sci. USA 80 (1983) 4403-4407] in the two-thirds N-terminal region. The cDNA clone specific for L30 protein (pRL30) has a 394-bp insert. The aa sequence of L30 protein was deduced from the nucleotide sequence of the cDNA. The protein consists of 114 aa residues with an Mr of 12 652. When compared with the N-terminal aa sequence of rat liver L30 protein [Wool, Annu. Rev. Biochem. 48 (1979) 719-754], pRL30 was found not to contain the initiation codon and 5'-noncoding region. The cDNA showed twelve silent changes in the coding region, one point mutation and one base deletion in the 3'-noncoding region, compared with mouse genomic DNA for L30 protein [Wiedemann and Perry, Mol. Cell Biol. 4 (1984) 2518-2528]. 相似文献
22.
Astrid Christiani Peter Hügelmeyer Prof. Dr. Wilhelm Stockem 《Cell and tissue research》1986,246(1):163-168
Summary Various stabilization and extraction procedures were tested to demonstrate the ultrastructural organization of the cytoskeleton in normal, locomoting Amoeba proteus. Most reliable results were obtained after careful fixation in glutaraldehyde/lysine followed by prolonged extraction in a polyethylene glycol/Triton X-100 solution. Before dehydration in a graded series of ethanol and critical-point drying, the amoebae were split by the sandwich-technique, i.e., by mechanical cleavage of cells mounted between two poly-L-lysine-coated glass slides. Platinum-carbon replicas as well as thin sections prepared from such cell fragments revealed a cytoskeleton composed of at least four different types of filaments: (1) 5–7-nm filaments organized as a more or less ordered cortical network at the internal face of the plasma membrane and probably representing F-actin; (2) 10–12-nm filaments running separately or slightly aggregated through the cytoplasm and probably representing intermediate filaments; (3) 24–26-nm filaments forming a loose network and probably representing microtubules; and (4) 2–4-nm filaments as connecting elements between the other cytoskeleton constituents. Whereas microfilaments are responsible for protoplasmic streaming and other motile phenomena, the function of intermediate filaments and cytoplasmic microtubules in amoebae is still obscure. 相似文献
23.
E Gianazza G Dossi F Celentano P G Righetti 《Journal of biochemical and biophysical methods》1983,8(2):109-133
A new technique for generating extended pH gradients (5 pH units) in Immobiline gels is reported. The previously described (J. Biochem. Biophys. Methods 7, 1983, 123-142) five-chamber gradient mixer has been replaced by a two-vessel device. A single mixture of the available Immobilines (pK 3.6, 4.6, 6.2, 7.0, 8.5 and 9.3) is made, with relative concentrations adjusted so as to produce the most uniform buffering power throughout the desired pH interval. This mixture is then divided into two portions, which are titrated to the extremes of the required pH span with an acidic titrant (Immobiline pK approximately 1) and a basic species (Immobiline pK 9.95). Highly reproducible pH gradients (pH 4-9) are thus generated, which appear extremely useful for the first dimensioned of 2-dimensional techniques. Our previously reported computer program has been implemented with an optimization algorithm which, given any cocktail of Immobilines, automatically adjusts the relative initial concentrations until the smoothest possible beta power is found. For the first time it is possible to perform IEF under controlled physico-chemical parameters: pH span and linearity, beta power, ionic strength and molarity of the buffering species. 相似文献
24.
25.
Neurotransmitter-Mediated Regulation of Brain Aromatase: Protein Kinase C- and G-Dependent Induction
Abstract: Aromatase in the diencephalic neurons, the level of which increases transiently during the prenatal to neonatal period, has been suggested to be involved in control of sexual behavior and differentiation of the CNS. Effects of neurotransmitters on levels of aromatase mRNA in cultured neurons were investigated to determine factors regulating the developmental increase that occurs in level of fetal brain aromatase. The expression of aromatase in diencephalic neurons of fetal mice at embryonic day 13, cultured in vitro, was significantly affected by α1 -adrenergic receptor ligands. Aromatase mRNA levels were higher in neurons treated with the α1 -agonist phenylephrine than in control neurons, whereas prazosin, an α1 -antagonist, suppressed this increase, and ligands for α2 - or β-adrenergic receptors did not exert any influence. The profile of α1 -adrenergic receptor subtypes during actual development in vivo suggested that the α1B subtype is in fact responsible for the signal transduction. Substance P, cholecystokinin, neurotensin, and brain natriuretic peptide also increased the level of expression along with phorbol 12-myristate 13-acetate and dibutyryl-cyclic GMP, whereas forskolin and dibutyryl-cyclic AMP caused a decrease. These data indicate that stimulation via α1 (possibly α1B )-adrenergic receptors, as well as receptors of specific neuropeptides, controls the expression of aromatase in embryonic day 13 diencephalic neurons through activation of protein kinase C or G. β-Adrenergic receptors would not appear to participate in the regulation, judging from their developmental profile, although cyclic AMP might be a suppressive second messenger. 相似文献
26.
Free Air Temperature Increase (FATI): a new tool to study global warming effects on plants in the field 总被引:4,自引:1,他引:3
I. NIJS F. KOCKELBERGH H. TEUGHELS H. BLUM G. HENDREY I. IMPENS 《Plant, cell & environment》1996,19(4):495-502
A new technique, called Free Air Temperature Increase (FATI), was developed to artificially induce increased canopy temperature in field conditions without the use of enclosures. This acronym was chosen in analogy with FACE (Free Air CO2 Enrichment), a technique which produces elevated CO2 concentrations [CO2] in open field conditions. The FATI system simulates global warming in small ecosystems of limited height, using infrared heaters from which all radiation below 800 nm is removed by selective cut-off filters to avoid undesirable photomorpho-genetic effects. An electronic control circuit tracks the ambient canopy temperature in an unheated reference plot with thermocouples, and modulates the radiant energy from the lamps to produce a 2.5°C increment in the canopy temperature of an associated heated plot (continuously day and night). This pre-set target differential is relatively-constant over time due to the fast response of the lamps and the use of a proportional action controller (the standard deviation of this increment was <1°C in a 3 week field study with 1007 measurements). Furthermore, the increase in leaf temperature does not depend on the vertical position within the canopy or on the height of the stand. Possible applications and alternative designs are discussed. 相似文献
27.
Grain yield, symbiotic N2 fixation and interspecific competition for inorganic N in pea-barley intercrops 总被引:4,自引:0,他引:4
Erik Steen Jensen 《Plant and Soil》1996,182(1):25-38
The effect of mixed intercropping of field pea (Pisum sativum L.) and spring barley (Hordeum vulgare L.), compared to monocrop cultivation, on the yield and crop-N dynamics was studied in a 4-yr field experiment using 15N-isotope dilution technique. Crops were grown with or without the supply of 5 g 15N-labeled N m-2. The effect of intercropping on the dry matter and N yields, competition for inorganic N among the intercrop components, symbiotic fixation in pea and N transfer from pea to barley were determined. As an average of four years the grain yields were similar in monocropped pea, monocropped and fertilized barley and the intercrop without N fertilizer supply. Nitrogen fertilization did not influence the intercrop yield, but decreased the proportion of pea in the yield. Relative yield totals (RYT) showed that the environmental sources for plant growth were used from 12 to 31% more efficiently by the intercrop than by the monocrops, and N fertilization decreased RYT-values. Intercrop yields were less stable than monocrop barley yields, but more stable than the yield of monocropped pea. Barley competed strongly for soil and fertilizer N in the intercrop, and was up to 30 times more competitive than pea for inorganic N. Consequently, barley obtained a more than proportionate share of the inorganic N in the intercrop. At maturity the total recovery of fertilizer N was not significantly different between crops, averaging 65% of the supplied N. The fertilizer N recovered in pea constituted only 9% of total fertilizer-N recovery in the intercrop. The amount of symbiotic N2 fixation in the intercrop was less than expected from its composition and the fixation in monocrop. This indicates that the competition from barley had a negative effect on the fixation, perhaps via shading. At maturity, the average amount of N2 fixation was 17.7 g N m-2 in the monocrop and 5.1 g N m-2 in the intercropped pea. A higher proportion of total N in pea was derived from N2 fixation in the intercrop than in the monocrop, on average 82% and 62%, respectively. The 15N enrichment of intercropped barley tended to be slightly lower than of monocropped barley, although not significantly. Consequently, there was no evidence for pea N being transferred to barley. The intercropping advantage in the pea-barley intercrop is mainly due to the complimentary use of soil inorganic and atmospheric N sources by the intercrop components, resulting in reduced competition for inorganic N, rather than a facilitative effect, in which symbiotically fixed N2 is made available to barley.Abbreviations MC
monocrop
- IC
intercrop
- PMC
pea monocrop
- BMC
barley monocrop
- PIC
pea in intercrop
- BIC
barley in intercrop 相似文献
28.
Ronald J. Prokopy Ratana Poramarcom Manon Sutantawong Ranu Dokmaihom Jorge Hendrichs 《Journal of Insect Behavior》1996,9(1):133-142
Oriental fruit flies,Bactrocera dorsalis, cultured in the laboratory for six generations, were released when 12–14 days old in an orchard of nonfruiting host trees that were furnished with either food and water, nonpunctured host fruit, punctured host fruit, or no resources. Nearly all flies of both sexes, all sexual behavior of males, and all mating pairs were observed on trees with fruit. Moreover, on trees with fruit, nearly all flies of both sexes, nearly all sexual behavior of males, and nearly all mating pairs occurred on the fruit itself rather than on the foliage or branches. In a subsequent test, both sexes were found to be strongly attracted to the odor of host fruit. For a polyphagous species of a tropical tephritid, these findings are the first to show a high level of male aggregation and a high amount of male sexual behavior on the fruit of host trees. Findings are discussed in relation to current knowledge of tephritid mating behavior. An additional quality-control test for laboratory cultured males used in the sterile insect technique of fly management is recommended. 相似文献
29.
B. E. Nixdorf-Bergweiler E. Wallhusser-Franke T. J. Devoogd 《Developmental neurobiology》1995,27(2):204-215
We investigated the development of spiny neurons in the lateral magnocellular nucleus of the anterior neostriatum before, during, and after song learning in male zebra finches (Taeniopygia guttata). The frequency of dendritic spines, dendritic field size, and branching characteristics were quantified at different ages in Golgi-stained tissue using a three-dimensional computerized tracing system. During development, overall spine frequencies increase between 3 and 5 weeks and decrease thereafter. In particular, spine frequencies of middle segments decrease significantly by 14% between 5 and 7 weeks posthatching (p = 0.017). A further reduction of 48% occurs between 7 weeks and adulthood (p < 0.001), resulting in a spine reduction of 56% on middle segments between 35 days of age and adulthood. In addition to the reduction of spine frequencies, we find regressive events also on some of the neuronal parameters that we have quantified. In general, dendrites of adult animals terminate closer to the cell body than those of 7-, 5-, or 3-week-old birds. Whereas no changes in segment length of first- and second-order dendrites have been identified, third-order dendrites end 19% closer to the cell body in adults than in younger birds (p < 0.024). Second-order dendrites in adult animals branch less frequently than in 3-week-old animals (35%, p = 0.017). There is also a trend of a smaller number of tertiary branches in adulthood compared with 3-week-old birds (41%, p = 0.060). The morphological changes may be related to the function of this nucleus and the sensitive phase for song acquisition. © 1995 John Wiley & Sons, Inc. 相似文献
30.
Hydrochlorothiazide (HCTZ) was shown to inhibit the transepithelial NaCl transport and the apical Na+-Cl? symport and to depolarize the apical membrane potential in the rabbit gallbladder epithelium. The depolarization was likely related to the opening of a Cl? conductance. To better understand whether an apical Cl? leak is involved in the mechanism of action of HCTZ, the transapical Cl? backflux was measured radiochemically by the washout technique. The gallbladder wall, pretreated with pronase on the serosal side to homogenize the subepithelium, was loaded with 36Cl? on the luminal side; mucosal and serosal 36Cl? effluxes (J m , J s ) were then measured every 2 min. The pretreatment with pronase did not alter the membrane potentials and the selectivity of the epithelium. Under control conditions and the tissue in steady-state, J m and J s time courses were each described by two exponential decays (A,B); the rate constants, k A and k B , were 0.71 ±0.03 and 0.16±0.01 min?1, respectively, and correspondingly the half-times (t 1 2A , t 1 2B ) were 1.01±0.05 and 5.00±0.44 min (n=10); these parameters were not significantly different for J m and J s time courses. J s was always greater than J m (J s /J m =2.02±0.22 and 1.43 ±0.17 for A and B decays). Under SCN? treatment in steady-state conditions, both J m and J s time courses were described by only one exponential decay, the component B being abolished. Moreover t 1 2A was similar to that predictable for the subepithelium. It follows that it is the component B which exits the epithelial compartment. Based on the intracellular specific activity and 36Cl? J m B at 0 min time of the washout experiment, the cell-lumen Cl? backflux in steady-state was calculated to be equal to about 2 μmol cm?2hr?1, in agreement with the value indirectly computable by other techniques. The experimental model was well responsive to different external challenges (increases in media osmolalities; luminal treatment with nystatin). HCTZ (2.5 · 10?4 m) largely increased 36Cl? J m B . The increase was abolished by luminal treatment with 10?4 m SITS, which not only brought back the efflux time courses to the ones observed under control conditions but even increased J s /J m of the cellular component, an indication of a reduced J m B . It is concluded that HCTZ opens an apical, SITS-sensitive Cl? leak, which contributes to dissipate the intracellular Cl? accumulation and to inhibit the NaCl transepithelial transport. Moreover, the drug is likely to reduce the basal electroneutral Cl? backflux supported by Na+-Cl? cotransport, in agreement with the inhibition of the cotransport itself. 相似文献