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41.
Dominique Rumeau Stephan Cuiné Laurent Fina Nathalie Gault Michel Nicole Gilles Peltier 《Planta》1996,199(1):79-88
The intracellular compartmentation of carbonic anhydrase (CA; EC 4.2.1.1), an enzyme that catalyses the reversible hydration of CO2 to bicarbonate, has been investigated in potato (Solanum tuberosum L.) leaves. Although enzyme activity was mainly located in chloroplasts (87% of total cellular activity), significant activity (13%) was also found in the cytosol. The corresponding CA isoforms were purified either from chloroplasts or crude leaf extracts, respectively. The cytosolic isoenzyme has a molecular mass of 255 000 and is composed of eight identical subunits with an estimated M
r of 30000. The chloroplastic isoenzyme (M
r 220000) is also an octamer composed of two different subunits with M
r estimated at 27 000 and 27 500, respectively. The N-terminal amino acid sequences of both chloroplastic CA subunits demonstrated that they were identical except that the M
r-27 000 subunit was three amino acids shorter than that of the M
r-27 500 subunit. Cytosolic and chloroplastic CA isoenzymes were found to be similarly inhibited by monovalent anions (Cl–, I–, N
3
-
and NO
3
-
) and by sulfonamides (ethoxyzolamide and acetozolamide). Both CA isoforms were found to be dependent on a reducing agent such as cysteine or dithiothreitol in order to retain the catalytic activity, but 2-mercaptoethanol was found to be a potent inhibitor. A polyclonal antibody directed against a synthetic peptide corresponding to the N-terminal amino acid sequence of the chloroplastic CA monomers also recognized the cytosolic CA isoform. This antibody was used for immunocytolocalization experiments which confirmed the intracellular compartmentation of CA: within chloroplasts, CA is restricted to the stroma and appears randomly distributed in the cytosol.Abbreviations BSA
bovine serum albumin
- CA
carbonic anhydrase
- PMSF
phenylmethylsulphonyl fluoride
- BAM
benzamidine
- DTT
dithiothreitol
- 2-ME
2-mercaptoethanol
- PVDF
polyvinylidene difluoride
The authors thanks P. Carrier and Dr. B. Dimon for technical assistance with the mass-spectrometry measurements. 相似文献
42.
Clare Gough Pascale Hemon Maurice Tronchet Christophe Lacomme Yves Marco Dominique Roby 《Molecular genetics and genomics : MGG》1995,247(3):323-337
A family of genes, the so-called msr genes (multiple stimulus response), has recently been identified on the basis of sequence homology in various plant species. Members of this gene family are thought to be regulated by a number of environmental or developmental stimuli, although it is not known whether any one member responds more specifically to one stimulus, or whether each gene member responds to various environmental stimuli. In this report, we address this question by studying the tobacco msr gene str246C. Using transgenic tobacco plants containing 2.1 kb of 5′ flanking DNA sequence from the str246C gene fused to the β-glucuronidase (GUS) coding region, the complex expression pattern of the str246C promoter has been characterized. Expression of the str246C promoter is strongly and rapidly induced by bacterial, fungal and viral infection and this induction is systemic. Elicitor preparations from phytopathogenic bacteria and fungi activate the str246C promoter to high levels, as do wounding, the application of auxin, auxin and cytokinin, salicylic acid or copper sulfate, indicating the absence of gene specialization within the msr gene family, at least for str246C. In addition, GUS activity was visualized. histochemically in root meristematic tissues of tobacco seedlings and is restricted to roots and sepals of mature plants. Finally, analysis of a series of 5′ deletions of the str246C promoter-GUS gene fusion in transgenic tobacco plants confirms the involvement of multiple regulatory elements. A region of 83 by was found to be necessary for induction of promoter activity in response to Pseudomonas solanacearum, while auxin inducibility and root expression are apparently not controlled by this element, since its removal does not abolish either response. An element of the promoter with a negative effect on promoter activation by P. solanacearum was also identified. 相似文献
43.
Eric Dufour Paul Robert Dominique Bertrand Tomasz Haertlé 《Journal of Protein Chemistry》1994,13(2):143-149
Fourier transform infrared spectroscopy has been applied to investigate the secondary structural changes of-lactoglobulin in water/ethanol mixtures. The studies were carried out at two differentpHs and at high protein concentrations. The spectra were recorded using an attenuated total reflection cell. The amide I band of-lactoglobulin in water reveals large amounts of intra extended-sheet structure. About 20% ethanol,-lactoglobulin unfolds and-strand formation is observed.-Helices are built up by increasing the ethanol concentration up to 30%. In 50% ethanol,-lactoglobulin gels providing the apparent pH are neutral. The secondary structural changes of-lactoglobulin were observed on the similarity maps obtained by Principal Component Analysis. 相似文献
44.
Virginia Avellana-Adalid Gerard Rebel Michel Caron Jean-Denis Cornillot Dominique Bladier Raymonde Joubert-Caron 《Glycoconjugate journal》1994,11(4):286-291
The distribution of a 14.4 kDa S-type lectin was examined in murine neuroblastoma cells, either undifferentiated or after differentiation induced by dibutyryl-cyclic adenosine monophosphate. In undifferentiated cells the immunoreactivity was detected extracellularly, associated with the plasma membrane and in bulges released into the extracellular milieu. Important modifications of the lectin localization were associated with the differentiation process that induced an increased cytosolic expression and a decreased externalization. Possible functions for the lectin expressed intracellularly in the differentiated cells are also considered. 相似文献
45.
Peter Giesbrecht Thomas Kersten Heinrich Maidhof Dominique Krüger Peter Blümel Harald Grob Jörg Wecke 《Archives of microbiology》1994,161(5):370-383
In log-phase cells of staphylococci, cultivated under high, non-lytic concentrations of penicillin G, there occurred a novel killing process hitherto hidden behind seemingly bacteriostatic effects. Two events are essential for the apprearance of this hidden death: (i) the failure of the dividing cell to deposit enough fibrillar cross-wall material to be welded together, and (ii) a premature ripping up of incomplete cross walls along their splitting system. Hidden death started as early as 10–15 min after drug addition, already during the first division cycle. It was the consequence of a loss of cytoplasmic constituents which erupted through peripheral slit-like openings in the incomplete cross walls. The loss resulted either in more or less empty cells or in cell shrinkage. These destructions could be prevented by raising the external osmotic pressure. In contrast, the conventional non-hidden death occurred only much later and exclusively during the second division cycle and mainly in those dividing cells, whose nascent cross walls of the first division plane had been welded together. These welding processes at nascent cross walls, resulting in tough connecting bridges between presumptive individual cells, were considered as a morphogenetic tool which protects the cells, so that they can resist the otherwise fatal penicillin-induced damages for at least an additional generation time (morphogenetic resistance system). Such welded cells, in the virtual absence of underlying cross-wall material, lost cytoplasm and were killed via ejection through pore-like wall openings or via explosions in the second division plane and after liberation of their murosomes, as it was the case in the presence of low, lytic concentrations of penicillin. Bacteriolysis did not cause any of the hitherto known penicillin-induced killing processes.Dedicated to Prof. Dr. Georg Henneberg on the occasion of his 85th birthday 相似文献
46.
Luc Malaval Dominique Modrowski Ashwani K. Gupta Jane E. Aubin 《Journal of cellular physiology》1994,158(3):555-572
The regulation of cell surface fibroblast growth factor (FGF) receptors during the differentiation of F9 teratocarcinoma cells was investigated. The capacity of F9 cells to bind 125I-basic FGF (FGF-2) increased upon induction of differentiation with dibutyryl cAMP and retinoic acid. No change in binding capacity was observed in the first 24 h after addition of differentiating agents, but a sixfold increase in binding capacity was observed after 48 h and a fivefold increase after 72 h. Scatchard analysis of the binding data indicated that the increased binding of 125I-FGF-2 was due to an increase in the number of receptors with no change in their affinity. When 125I-FGF-2 was cross-linked to cell surface receptors, an increase in FGF-2-receptor complexes with molecular weights of 140,000–160,000 was also observed in the differentiated F9 cells. Undifferentiated F9 cells are known to secrete FGF-4 and cease expression of this molecule upon differentiation. To determine whether the low level of receptors in undifferentiated cells might be related to their production of FGF ligands, the ability of suramin, a drug that can disrupt FGF-receptor interactions, to modulate receptor number on F9 cells was investigated. Suramin treatment increased 125I-FGF-2 binding capacity of undifferentiated F9 cells threefold but had little effect on the binding capacity of differentiated cells. In addition, antibodies to FGF-4 increased the 125I-FGF-2 binding capacity of undifferentiated F9 cells by 58%. These results suggest that undifferentiated F9 cells might be responding in an autocrine manner to their own FGF ligands resulting in downregulation of cell surface FGF receptors. The increased number of receptors observed in differentiated cells may partly result from the decreased production of FGF ligands by these cells. © 1994 Wiley-Liss, Inc. 相似文献
47.
48.
Cysteine proteinase forms in sprouting potato tuber 总被引:1,自引:0,他引:1
Dominique Michaud Binh Nguyen-Quoc Nathalie Bernier-Vadnais Loïc Faye Serge Yelle 《Physiologia plantarum》1994,90(3):497-503
Transformation of plants with exogenous proteinase inhibitor genes represents an attractive strategy for the biological control of insect pests. However, such a strategy necessitates a thorough characterization of endogenous proteinases. which represent potential target enzymes for the exogenous inhibitors produced. In the present study. changes in general endoproteolytic activity were monitored during sprouting of potato ( Solanum tuberosum L. cv. Kennebec) tuber. Quantitative data obtained using standard procedures showed that an increase in cysteine proteinase (EC 3.4.22) activity occurs during sprouting. This increased activity results from the gradual appearance of new cysteine proteinase forms, as demonstrated by the use of class-specific proteinase activity gels. While only one cysteine proteinase form was present during early sprouting, at least six new active forms of the same class were shown to appear gradually after the mature tuber was sown, suggesting the involvement of a complex cysteine proteolytic system in the last stages of tuber protein breakdown. Interestingly, oryzacystatins I and II. two cysteine proleinase inhibitors potentially useful for insect control, had no effect on any tuber proteinase delected. Similar results were obtained with leaf, stem and stolon proteinases. This apparent absence of direct interference supports the potential of oryzacystatin genes for production of insect-tolerant transgenie potato plants. 相似文献
49.
Changes in carbon storage in temperate humic loamy soils after forest clearing and continuous corn cropping in France 总被引:3,自引:0,他引:3
Soil samples from forest and agricultural sites in three areas of southwest France were collected to determine the effect
of forest conversion to continuous intensive corn cropping with no organic matter management on soil organic carbon (C) content.
Soils were humic loamy soils and site characteristics that may affect soil C were as uniform as possible (slope, elevation,
texture, soil type, vegetation).
Three areas were selected, with adjacent sites of various ages of cultivation (3 to 35 yr), and paired control forest sites.
The ploughed horizon (0-Dt cm) and the Dt-50 cm layer were collected at each agricultural site. In forest sites, each 10 cm
layer was collected systematically down to 1 meter depth. Carbon concentrations were converted to total content to a given
depth as the product of concentration, depth of sample and bulk density, and expressed in units of kg m-2. For each site and each sampled layer, the mineral mass of soil was calculated, in order to base comparisons on the same
soil mass rather than the same depth.
The pattern of C accumulation in forest soils showed an exponential decrease with depth. Results suggested that soil organic
carbon declined rapidly during the first years of cultivation, and at a slower rate thereafter. This pattern of decrease can
be fitted by a bi-exponential model assuming that initial soil organic carbon can be separated into two parts, a very labile
pool reduced during the first rapid decline and more refractory fractions oxidizing at a slower rate. Sampling to shallow
depths (0-Dt cm) resulted in over-estimation of the rate of carbon release in proportion to the initial amount of C, and in
under-estimation of the total loss of C with age. The results for the 0–50 cm horizon indicated that losses of total carbon
average about 50% in these soils, ranging in initial carbon content from 19 to 32.5 kg m-2. Carbon release to the atmosphere averaged 0.8 kg m-2 yr-1 to 50 cm depth during the first 10 years of cultivation. The results demonstrate that temperate soils may also be an important
source of atmospheric carbon, when they are initially high in carbon content and then cultivated intensively with no organic
matter management. 相似文献
50.
Bienvenu Thierry Hubert Dominique Fonknechten Nuria Dusser Daniel Kaplan Jean Claude Beldjord Cherif 《Human genetics》1994,94(1):65-68
Human Genetics - Cystic fibrosis is caused by mutations in the cystic fibrosis transmembrane conductance regulator gene (CFTR). Analysis of DNA from a pancreatic sufficient patient by means of... 相似文献