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101.
Agrobacterium tumefaciens-mediated barley transformation 总被引:26,自引:2,他引:24
Sonia Tingay David McElroy Roger Kalla Sarah Fieg Mingbo Wang Sarah Thornton Richard Brettell 《The Plant journal : for cell and molecular biology》1997,11(6):1369-1376
Genetically transformed barley was produced by eco-cultivating immature embryo explants with Agrobacterium tumefaciens carrying a binary vector coding for chimaeric bacterial genes, bar and gus , and selecting for bialaphos-resistant cultures from which plants were regenerated. Integration of both genes was confirmed by gel blot hybridization analysis of DNA from the transformed plants and their progenies. From 1282 embryos, plants were recovered for 54 independently transformed lines, giving a transformation efficiency of 4.2%. Transgene numbers in the different lines ranged from single copy insertion to at least ten copies. Sixteen out of 18 plants grown to maturity were fully fertile. Both marker genes, bar and gus , were expressed and co-segregated in the T1 progeny plants. In the majority of cases, the genes showed Mendelian segregation predicted for transgene insertion at a single locus. In one family with multiple transgene insertions, molecular analysis of T1 and T2 plants suggested that the T-DNA had inserted at two unlinked loci. 相似文献
102.
A defect in synapsis causes male sterility in a T-DNA-tagged Arabidopsis thaliana mutant 总被引:4,自引:2,他引:2
Brenda N. Peirson Sarah E. Bowling Christopher A. Makaroff 《The Plant journal : for cell and molecular biology》1997,11(4):659-669
Fluorescence microscopy was used to study meiosis in microsporocytes from wild-type Arabidopsis thaliana and a T-DNA-tagged meiotic mutant. Techniques for visualizing chromosomes and β-tubulin in other plant species were evaluated and modified in order to develop a method for analyzing meiosis in A. thaliana anthers. Like most dicots, A. thaliana microsporocytes undergo simultaneous cytokinesis in which both meiotic divisions are completed prior to cytokinesis. However, two unique events were observed in wild-type A. thaliana that have not been reported in other angiosperms: (1) polarization of the microsporocyte cytoskeleton during prophase I prior to nuclear envelope breakdown, and (2) extensive depolymerization of microtubules just prior to metaphase II. The first observation could have implications regarding a previously uncharacterized mechanism for determining the axis of the metaphase I spindle during microsporogenesis. The second observation is peculiar since microtubules are known to be involved in chromosome alignment in other species; possible explanations will be discussed. A T-DNA-tagged meiotic mutant of A. thaliana ( syn1 ), which had previously been shown to produce abnormal microspores with variable DNA content, was also cytologically characterized. The first observable defect occurs in microsporocytes at telophase I, where some chromosomes are scattered throughout the cytoplasm, usually attached to stray microtubules. Subsequent developmental stages are affected, leading to complete male sterility. Based on similarities to synaptic mutants that have been described in other species, it is suggested that this mutant is defective in synaptonemal complex formation and/or cohesion between sister chromatids. 相似文献
103.
104.
M. T. Gonzlez-Muoz M. L. Merroun N. Ben Omar J. M. Arias 《International biodeterioration & biodegradation》1997,40(2-4)
This paper deals with uranium biosorption by Myxococcus xanthus biomass in which dry biomass, accumulating up to 2.4 mM of uranium g−1, is demonstrated to be a more efficient biosorbent than wet biomass. For uranium concentrations of 0.1–0.3 mM, between 95.79% and 95.99% of the uranium was taken up from the solution. Dry biomass biosorption was found to be relatively rapid, reaching equilibrium after 5–10 min. In addition, the pH influenced biosorption, pH 4.5 promoting maximum uptake. It was also established that the biosorbed uranium is located on the cellular wall and within the extracellular mucopolysaccharide of this microorganism. Furthermore, using sodium carbonate as a desorbent agent, 80.82% of the biosorbed uranium could be recovered. The results obtained indicate the possible utilization of M. xanthus biomass to solve some problems of the water contaminated by uranium. 相似文献
105.
P-Element Insertion Alleles of Essential Genes on the Third Chromosome of Drosophila Melanogaster: Correlation of Physical and Cytogenetic Maps in Chromosomal Region 86e-87f 总被引:2,自引:0,他引:2
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P. Deak M. M. Omar RDC. Saunders M. Pal O. Komonyi J. Szidonya P. Maroy Y. Zhang M. Ashburner P. Benos C. Savakis I. Siden-Kiamos C. Louis V. N. Bolshakov F. C. Kafatos E. Madueno J. Modolell D. M. Glover 《Genetics》1997,147(4):1697-1722
We have established a collection of 2460 lethal or semi-lethal mutant lines using a procedure thought to insert single P elements into vital genes on the third chromosome of Drosophila melanogaster. More than 1200 randomly selected lines were examined by in situ hybridization and 90% found to contain single insertions at sites that mark 89% of all lettered subdivisions of the Bridges' map. A set of chromosomal deficiencies that collectively uncover ~25% of the euchromatin of chromosome 3 reveal lethal mutations in 468 lines corresponding to 145 complementation groups. We undertook a detailed analysis of the cytogenetic interval 86E-87F and identified 87 P-element-induced mutations falling into 38 complementation groups, 16 of which correspond to previously known genes. Twenty-one of these 38 complementation groups have at least one allele that has a P-element insertion at a position consistent with the cytogenetics of the locus. We have rescued P elements and flanking chromosomal sequences from the 86E-87F region in 35 lines with either lethal or genetically silent P insertions, and used these as probes to identify cosmids and P1 clones from the Drosophila genome projects. This has tied together the physical and genetic maps and has linked 44 previously identified cosmid contigs into seven ``supercontigs' that span the interval. STS data for sequences flanking one side of the P-element insertions in 49 lines has identified insertions in the αγ element at 87C, two known transposable elements, and the open reading frames of seven putative single copy genes. These correspond to five known genes in this interval, and two genes identified by the homology of their predicted products to known proteins from other organisms. 相似文献
106.
Roles for Protein Kinase C and Mitogen-Activated Protein Kinase in Nicotine-Induced Secretion from Bovine Adrenal Chromaffin Cells 总被引:3,自引:2,他引:1
Abstract: Both the Ca2+/phospholipid-dependent protein kinases (protein kinases C, PKCs) and mitogen-activated protein kinases (MAPKs) have been implicated as participants in the secretory response of bovine adrenomedullary chromaffin cells. To investigate a possible role for these kinases in exocytosis and the relationship of these kinases to one another, intact chromaffin cells were treated with agents that inhibited each of the kinases and analyzed for catecholamine release and MAPK/extracellular signal-regulated kinase (ERK) kinase (MEK)/MAPK activation after stimulation with secretagogues of differential efficacy. Of the three secretagogues tested, inactivation of PKCs by long-term phorbol 12-myristate 13-acetate (PMA) treatment or incubation with GF109203X had the greatest inhibitory effect on nicotine-induced catecholamine release and MEK/MAPK activation, a moderate effect on KCl-induced events, and little, if any, effect on Ca2+ ionophore-elicited exocytosis and MEK/MAPK activation. These results indicate that PKC plays a significant role in events induced by the optimal secretagogue nicotine and a lesser role in exocytosis elicited by the suboptimal secretagogues KCl and Ca2+ ionophore. Treatment of cells with the MEK-activation inhibitor PD098059 completely inhibited MEK/MAPK activation (IC50 1–5 µM) and partially inhibited catecholamine release induced by all secretagogues. However, PD098059 was more effective at inhibiting exocytosis induced by suboptimal secretagogues (IC50~10 µM) than that induced by nicotine (IC50~30 µM). These results suggest a more prominent role for MEK/MAPK in basic secretory events activated by suboptimal secretagogues than in those activated by the optimal secretagogue nicotine. However, PD098059 also partially blocked secretion potentiated by short-term PMA treatment, suggesting that PKC can function in part by signaling through MEK/MAPK to enhance secretion. Taken together, these results provide evidence for the preferential involvement of MEK/MAPK in basic secretory events activated by the suboptimal secretagogues KCl and Ca2+ ionophore and the participation of both PKC and MEK/MAPK in optimal secretion induced by nicotine. 相似文献
107.
Omar Benzakour Chryso Kanthou Florea Lupu Ulla Dennehy Chris Goodwin Michael F. Scully Vijay V. Kakkar David N. Cooper 《Journal of cellular biochemistry》1995,59(4):514-528
Thrombin is a potent mitogen for human vascular smooth muscle cells (HVSMC) and its enzymatic activity is required for this function. The present study demonstrates that prothrombin is also mitogenic for HVSMC due to the generation of enzymatically active thrombin which occurs upon incubation of prothrombin with the cells. Analysis by SDS-PAGE, immunoblotting, and amino acid sequencing revealed that prothrombin incubated with HVSMC undergoes limited proteolysis. Prethrombin 1 was formed through cleavage at R155-S156. Cleavage at R271-T272 generated fragment 1.2 and prethrombin 2 whilst cleavage at R284-T285 yielded truncated prothrombin 2 (prethrombin 2′). However, cleavage at R320-I321 which, during prothrombin activation produces two-chain α-thrombin, was not detectable. Studies on HVSMC-conditioned medium revealed that a similar pattern of prothrombin cleavage occurred by a cell-secreted factor(s). Amidolytic activity analysis indicated that 1–3% catalytically active thrombin-like activity was generated upon incubation of prothrombin with HVSMC-conditioned medium. By treating conditioned medium with various classes of proteinase inhibitors or hirudin, it was determined that prothrombin is cleaved by a cell-derived serine proteinase-like factor(s) at R271-S272 and by α-thrombin at R155-S156 and R284-T285. Antibodies neutralising the activity of either urokinase, tissue plasminogen activator, or factor Xa failed to alter the prothrombin cleaving activity of conditioned medium. This activity which may catalyse an alternative pathway for the generation of thrombin, was eluted from a gel filtration column as a single peak with apparent molecular mass of 30–40 kDa. © 1995 Wiley-Liss, Inc. 相似文献
108.
Genetic regulation of gibberellin deactivation in Pisum 总被引:2,自引:0,他引:2
John J. Ross James B. Reid Stephen M. Swain Omar Hasan rew T. Poole Peter Hedden Christine L. Willis 《The Plant journal : for cell and molecular biology》1995,7(3):513-523
The regulation of gibberellin (GA) deactivation was examined using the sin (slender) mutation in the garden pea (Pisum sativum L.). This mutation blocks the deactivation of GA20, the precursor of the bioactive GA1. Firstly, crosses were made to combine sin with the GA biosynthesis mutations na, lhi and le-3. The combination sin na produced a novel phenotype, with long (‘slender’) basal internodes and extremely short (‘nana’) upper internodes. In contrast, the double mutant sin lhi was phenotypically dwarf. The mutation sin causes an accumulation of GA20 in maturing seeds, and this was unaffected by na, since the na mutation is not expressed in seeds. In contrast, lhi seeds did not accumulate GA20, since lhi imposes an early block on GA biosynthesis. Secondly, the effects of sin on several steps in GA deactivation were investigated. In maturing seeds, the mutation sin blocks two steps in GA20 metabolism, namely, GA20 to GA29, and GA29 to GA29-catabolite. In the vegetative plant, on the other hand, sin blocked the step GA20 to GA29, but not GA29 to GA29-catabolite; the steps GA20 to GA81 and GA20 to GA1 were also not impaired in this mutant. It is clear that the effects of sin, like those of na, are strongly organ-specific. The presence of separate enzymes for the steps GA20 to GA29 and GA29 to GA29-catabolite was suggested by the observation that GA8 inhibited the latter step, but not the former, and by the inability of GA20 and GA29 to inhibit each other's metabolism. It is suggested that the Sin gene may be a regulatory gene controlling the expression of two structural genes involved in GA deactivation. 相似文献
109.
Gibberellins A1, A4, A9, A12-aldehyde, A20 and A51, each labelled with both a radioactive and stable isotope were fed to immature barley grain by injection into the endosperm. After 7 d, extensive metabolism of all substrates had occurred, and metabolites were identified by combined capillary gas chromatography-mass spectrometry. A proposed scheme of gibberellin metabolism in immature barley grain is presented.Abbreviations GAn
gibberellin An
- GC-MS
combined gas chromatography-mass spectrometry
- HPLC
high-performance liquid chromatography 相似文献
110.
Ernesto Bonilla Enrique Salazar Jose Joaquin Villasmil Ruddy Villalobos Magaly Gonzalez Jose Omar Davila 《Neurochemical research》1984,9(11):1543-1548
Copper concentration was determined in samples from 38 areas of 7 normal human brains. The grey matter contained higher concentrations of copper than the white matter. Identical areas of the grey and white matter of the cerebral cortex showed significant differences between individuals. In the caudate nucleus the highest concentrations of copper were found in the tail followed by the body and the head, respectively. A negative linear regression between age and brain copper levels was demonstrated. 相似文献