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1.
Stimulation of poly(ADP-ribose) synthesis by free radicals in C3H10T1/2 cells: relationship with NAD metabolism and DNA breakage 总被引:1,自引:0,他引:1
D Lautier D Poirier A Boudreau M A Alaoui Jamali A Castonguay G Poirier 《Biochimie et biologie cellulaire》1990,68(3):602-608
We have studied the effect of H2O2 and O2- produced by xanthine and xanthine oxidase on NAD catabolism, poly(ADP-ribose) synthesis, and production of DNA single-strand breaks in C3H10T1/2 cells. The results show a correlation between the induction of DNA single-strand breaks, the decrease of NAD pool, and the accumulation of polymer. New techniques, based on affinity chromatography and reversed-phase high pressure liquid chromatography, have allowed an accurate determination of polymer contents and showed a 20-fold stimulation of polymer biosynthesis induced by active oxygen species. Inhibition experiments performed with 3-aminobenzamide have shown that the decrease in NAD levels after exposure of cells to active oxygen species was caused by stimulation of poly(ADP-ribosyl)ation and of another cellular process. 相似文献
2.
Electrophoretic studies of the degree and pattern of polymorphism at two third-chromosome loci, esterase-6 (Est-6) and phosphoglucomutase (PGM), were carried out in three Drosophila melanogaster populations collected from different localities in Iraq: Mosul, Tuwaitha, and Basrah. The results show that only the Tuwaitha population was polymorphic for both loci; the other two populations were polymorphic for Est-6 and monomorphic for PGM. The allele frequency changes at both loci were followed for 20 generations in an experimental cage derived from the Tuwaitha population; it was found that there is a deviation from Hardy-Weinberg equilibrium at both loci toward the homozygote. 相似文献
3.
J. Ballongue J. Amine E. Masion H. Petitdemange R. Gay 《FEMS microbiology letters》1985,29(3):273-277
Abstract Factors that may initiate the biosynthesis of acetoacetate decarboxylase were investigated in resting cells of Clostridium acetobutylicum . Linear acids from C1 to C4 were inducers, whereas branched acids and linear acids from C5 to C7 were not inducers of acetoacetate decarboxylase biosynthesis. Induction of acetoacetate decarboxylase was maximal at pH 4.8 in the presence of acid concentrations comparable with those found during fermentation. In growth conditions repression of acetoacetate decarboxylase biosynthesis was found. This fact explains that acetone production by Clostridium acetobutylicum occurs when growth slows down. 相似文献
4.
Scale-up of the adenovirus expression system for the production of recombinant protein in human 293S cells 总被引:6,自引:0,他引:6
Alain Garnier Johanne Côté Isabelle Nadeau Amine Kamen Bernard Massie 《Cytotechnology》1994,15(1-3):145-155
Human 293S cells, a cell line adapted to suspension culture, were grown to 5×106 cells/mL in batch with calcium-free DMEM. These cells, infected with new constructions of adenovirus vectors, yielded as much as 10 to 20% recombinant protein with respect to the total cellular protein content. Until recently, high specific productivity of recombinant protein was limited to low cell density infected cultures of no more than 5×105 cells/mL. In this paper, we show with a model protein, Protein Tyrosine Phosphatase 1C how high product yield can be maintained at high cell densities of 2×106 cells/mL by a medium replacement strategy. This allows the production of as much as 90 mg/L of active recombinant protein per culture volume. Analysis of key limiting/inhibiting medium components showed that glucose addition along with pH control can yield the same productivity as a medium replacement strategy at high cell density in calcium-free DMEM. Finally, the above results were reproduced in 3L bioreactor suspension culture thereby establishing the scalability of this expression system. The process we developed is used routinely with the same success for the production of various recombinant proteins and viruses.Abbreviations CFDMEM
calcium-free DMEM
- CS
bovine calf serum
- hpi
hours post-infection
- J+
enriched Joklik medium
- MLP
major late promoter
- MOI
multiplicity of infection (# of infectious viral particle/cell)
- q
specific consumption rate (mole/cell.h)
- pfu
plaque forming unit (# of infectious viral particle)
- Y
yield (g/E6 cells or mole/cell) 相似文献
5.
An in vitro root explant tissue culture technique is described for determining susceptibility of tomato (Lycopersicon esculentum Mill.) breeding lines and cultivars to the root-knot nematode Meloidogyne incognita. Root explants were taken from 2-day-old seedlings cultured for 30 days at 28 C on Gamborg''s B-5 medium with or without nematode inoculum. The remaining portion of the root and stem from the excised root explants was transferred to soil in pots and grown to maturity in the greenhouse. In vitro root explants were evaluated for growth and occurrence of juveniles, adults, and egg masses. The regenerated plants were used to produce more seed, The proposed technique is simple, reliable, and adapted to routine screening of large numbers of F₁ and F₂ samples, and it utilizes less space than tests performed on intact plants in the greenhouse or growth chamber. Evidence is presented also on the breakdown of resistance to M. incognita under high temperature stress using this in vitro root explant technique. 相似文献
6.
Phosphorus exchange at the sediment-water interface coupled with several parameters were assessed in several reservoirs with geologically different catchment basins and different trophic status in Morocco and France.The results showed that these exchanges were regulated by a combination of factors: physical chemical variability of the environment, the geological composition of catchment basins and the trophic status of the lake.In the hypereutrophic Villerest, iron-bound phosphorus is the major form of phosphorus trapped by the sediment whereas, in Moroccan reservoirs, calcium-bound phosphorus prevailed.We suggest that a drastic control of phosphorus inputs into the waters must be done through a large program of dephosphatization of tributaries to avoid Microcystis aeruginosa bloom formation in Villerest (Aleya et al., 1993) and calcium-bound phosphorus dissociation in Moroccan reservoirs with upward release of bioavailable phosphorus.
Résumé Les échanges de phosphore au niveau de l'interface eau-sédiment couplés á la distribution temporelle de divers éléments chimiques et biologiques ont été étudiés dans divers réservoirs de niveaux trophiques différents, au Maroc et en France.Nos résultats mettent clairement en évidence une influence directe de l'environnement physico-chimique, de la nature géologique des bassins versants et de l'état trophique du lac sur la dynamique du phosphore au sein de cette interface.De plus, il apparait que dans le lac hypereutrophe de Villerest (Roanne, France), le phosphore est majoritairement complexé au fer alors que dans les retenues marocaines, ce sont les complexes phosphore-calcium qui prédominent.Nous préconisons un contrôle drastique des apports en phosphore á travers l'installation et la multiplication d'unités de déphosphatation afin d'éviter d'une part, la prolifération massive de la Cyanobactérie Microcystis aeruginosa á Villetest (Aleya et al., 1994) et d'autre part la dissociation des complexes phhosphore-calcium au sein des retenues marocaines avec libération de phosphore biodisponible.相似文献
7.
Portulaca oleracea leaves were found to contain 0.42% of a mucilage mixture. The mucilage was fractionated into an acidic and a neutral fraction. The acidic fraction consists of galacturonic acid residues joined by α-(1→4)-linkages; 60% of these residues are present as the calcium salt, and esterified galacturonic acid residues are absent. The neutral fraction is composed of 41% of arabinose and 43% of galactose residues, besides traces of rhamnose residues. 相似文献
8.
Studies were undertaken on a highly metastatic hamster fibrosarcoma cell line with a view to assessing whether cells entering into apoptosis, measured by counting the number of transglutaminase mediated detergent insoluble envelopes, has any synchrony with a particular phase of the cell cycle. A double exposure of thymidine was used to block cells in early S-phase. Flow cytometry in combination with [3H]thymidine incorporation into DNA was used to assess the degree of synchrony and progression through the different phases of cell cycle. The apoptotic index was found to be at its maximum in mid-S-phase. Measurement of transglutaminase activity in each phase of the cell cycle indicated that the specific activity was also at its greatest during mid S-phase. The level of enzyme was relatively unchanged throughout the cell cycle indicating that the regulation of transglutaminase activity occurs primarily through effects on catalytic activity rather than enzyme synthesis. 相似文献
9.
Sanaa Hamdy Omar Siegfried Honecker Hans-Jürgen Rehm 《Applied microbiology and biotechnology》1992,36(4):518-524
Summary The formation of citric acid, oxalic acid, erythritol and glycerol by three strains of Aspergillus niger immobilized in calcium alginate was investigated and compared with that of free cells when cultivated in shake flasks under phosphate limitation. Morphological changes were followed using an electron microscope. The production of acids and polyols, the consumption of glucose and fructose, and also the morphological changes were strain-dependent. The results also reflected the influence of long storage of a strain on productivity, morphological behaviour and phosphate consumption.
Offprint requests to: H.-J. Rehm 相似文献
10.