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161.
Summary Induction of prophage occurs in recA441 mutant lysogens after a shift to 42° C in the presence of adenine. If the synthesis of RecA441 protein is maintained at a low basal level by the presence of a second mutation in the recA441 gene, recA453, induction of prophage is prevented. The ability to induce prophage is restored by the introduction, on a transducing phage, of a second recA gene carrying the recA430 mutation; by itself, the RecA430 protein is devoid of activity against the repressor (Rebollo et al. 1984). In order to explain how the RecA430 protein might complement the RecA441 protein to provide repressor cleavage in a recA453-441 (recA430) diploid lysogen, we characterized the cleavage reaction catalysed by a mixture of these proteins in vitro. Our results suggest that, in the presence of dATP, the RecA441 and RecA430 proteins form mixed multimers on single-stranded DNA, in which the RecA441 protein molecules enhance the DNA binding affinity of RecA430 protein molecules, but RecA430 protein molecules support no cleavage of the repressor.Although the effects of the RecA430 and single-strand binding (SSB) proteins are similar in vitro, we show that the SSB protein cannot substitute for the RecA430 protein in restoring repressor cleavage in a recA453-441 lysogen. Comparison of the stimulatory effect of long single-stranded DNA with that of (dA)14 oligonucleotides on the RecA441 protein-directed cleavage of the repressor in the presence of various nucleoside triphosphates (NTPs) indicates that the cooperative binding of the RecA441 protein to single-stranded DNA stabilizes the RecA protein-DNA complexes so that they remain intact long enough to support cleavage of the repressor. We conclude that the low basal level of the RecA441 protein in a recA453-441 cell is sufficient to cleave the repressor, under conditions where a normal basal level of RecA430 protein is also present allowing the formation of mixed multimers on single-stranded DNA regions normally present in the cell. 相似文献
162.
Ecdysteroid levels were determined during the period of the adult reproductive cycle in the ovovivparous fleshfly Sarcophaga bullata. Low levels were found in males during the 10 days following eclosion while the entire adult female showed a significant peak of ecdysteroid activity at 190 h post eclosion (i.e. during embryogenesis). When the female reproductive tract was analyzed it was observed that the ovaries became vitellogenic a short time after a protein meal was offered at 96 h post eclosion: at 140 h, ecdysteroid activity was recorded in the developing oöcytes. The major peak of ecdysteroids during the reproductive cycle was found in developing embryos at 190 h. The significance of these releases of ecdysteroids is discussed in relation to major embryonic developmental events. 相似文献
163.
Steven P. Butcher Peter J. Roberts James F. Collins† 《Journal of neurochemistry》1984,43(4):1039-1045
Abstract: The effects of ions on the binding of the excitatory amino acid analogue dl -[3 H]2-amino-4-phosphon-obutyrate to l -glutamate-sensitive sites on rat brain synaptic membranes was investigated. The divalent cations manganese, magnesium, strontium, and particularly calcium, produced a marked enhancement in specific binding. However, this effect was manifest only in the presence of added chloride, or to a lesser extent, with bromide ions. Application of saturation analysis revealed that both chloride and calcium acted to increase the binding site density in a concentration-dependent manner, without affecting the dissociation constant. The only other ionic species found to have a significant effect on 2-amino-4-phosphonobutyrate binding was sodium, which produced an apparent reduction in site affinity, without modifying the binding site density. Although the significance of these striking ionic effects is as yet unknown, it seems feasible that chloride (and possibly also calcium) ions may serve a role in regulating the interaction of excitatory amino acids with their physiological receptors. 相似文献
164.
165.
Corticotropin-releasing factor (CRF) stimulates locomotor activity in intact and hypophysectomized newts (Amphibia) 总被引:2,自引:0,他引:2
Locomotor activity of rough-skinned newts (Taricha granulosa) was significantly higher in intact and hypophysectomized males injected intracranially with 100 ng CRF (ovine corticotropin-releasing factor) than in those injected with 10 ng CRF or saline. In addition, an injection of corticosterone or dexamethasone failed to stimulate newt locomotor activity. These results provide evidence that CRF can act independently of pituitary hormones to stimulate locomotor activity in a nonmammalian vertebrate. 相似文献
166.
Identification of the DNA sequences encoding the large subunit of the mRNA-capping enzyme of vaccinia virus. 总被引:13,自引:5,他引:8
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The DNA sequences encoding the large subunit of the mRNA-capping enzyme of vaccinia virus were located on the viral genome. The formation of an enzyme-guanylate covalent intermediate labeled with [alpha-32P]GTP allowed the identification of the large subunit of the capping enzyme and was used to monitor the appearance of the enzyme during the infectious cycle. This assay confirmed that after vaccinia infection, a novel 84,000-molecular-weight polypeptide corresponding to the large subunit was rapidly synthesized before viral DNA replication. Hybrid-selected cell-free translation of early viral mRNA established that vaccinia virus encoded a polypeptide identical in molecular weight with the 32P-labeled 84,000-molecular-weight polypeptide found in vaccinia virions. Like the authentic capping enzyme, this virus-encoded cell-free translation product bound specifically to DNA-cellulose. A comparison of the partial proteolytic digestion fragments generated by V8 protease, chymotrypsin, and trypsin demonstrated that the 32P-labeled large subunit and the [35S]methionine-labeled cell-free translation product were identical. The mRNA encoding the large subunit of the capping enzyme was located 3.1 kilobase pairs to the left of the HindIII D restriction fragment of the vaccinia genome. Furthermore, the mRNA was determined to be 3.0 kilobases in size, and its 5' and 3' termini were precisely located by S1 nuclease analysis. 相似文献
167.
Anita K. Costa Dominic F. Heffel Thomas M. Schieble James R. Trudell 《In vitro cellular & developmental biology. Plant》1987,23(7):501-506
Summary Inasmuch as it is known that the toxicity of anesthetic agents is potentiated by hypoxia and that the reductive metabolism
of these agents results in the formation of lipid hydroperoxides, we investigated the toxicity of hydroperoxides under low-oxygen
concentrations. We found that hypoxia exacerbates the toxicity oft-butyl hydroperoxide, shifting the dose-response curve oft-butyl hydroperoxide vs. lysis of hepatocytes approximately an order of magnitude to the left. Furthermore, although at the
end of a 4-h exposure to 0.5% O2 hepatocyte monolayers seemed normal by three indices (release of51Cr and serum glutamate transaminase or exclusion of trypan blue), they were completely lysed after an additional 20 h reoxygenation
at 20%. O2. In contrast, monolayers exposed to 2% O2 for 4 h seemed normal after 20 h reoxygenation. However, cells exposed to both a subtoxic dose of hydroperoxide and 4 h of
2% O2, although seeming healthy at the end of the hypoxic period, were completely lysed within 20 h after reoxygenation.
The study was supported by grant OH 00978 from the National Institutes for Occupational Safety and Health, Atlanta, Georgia. 相似文献
168.
Marianne Klint Anita Fridberger Alan Menge Jan Sllstrm Leif Plen 《Molecular reproduction and development》1987,17(2):173-190
Boar sperm glycoprotein fractions were isolated by Lens culinaris hemagglutinin affinity chromatography of detergent-solubilized ejaculated spermatozoa, followed by preparative sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis. In order to develop methods for further investigations of the sperm proteins, we proceeded with two of the isolated glycoproteins. Antibodies were raised in female rabbits against each of the two sperm glycoproteins. By a combination of immunosorbent chromatography, using the antibodies obtained, and preparative SDS polyacrylamide gel electrophoresis, highly purified sperm proteins were isolated. The sperm proteins were immobilized on Sepharose gel columns and specific immunoglobulin Fab fragments were enriched by affinity chromatography. The specificity of the Fab fragments was ascertained by immunoprecipitation analysis. The Fab fragments were used in indirect immunofluorescence analysis to localize the corresponding antigens on the surface of boar spermatozoa. Both antigens were exclusively confined to the postacrosomal region. Immunohistochemical staining of boar testis sections revealed that both antigens are expressed from the spermatid stage. This technique also revealed that one of the antigens congregated at the Golgi complex-acrosome region during spermatogenesis. 相似文献
169.
The effect of bovine conceptus secretory proteins (CSP) on uterine prostaglandin (PG)-F2 alpha production was evaluated in dairy cattle following injection of estradiol-17 beta. Intrauterine injections of dialyzed serum proteins (Control, n = 5) or CSP (n = 5) were administered from days 15 through 18 post-estrus. Following intrauterine treatments on day 18, all cows were injected with E2 (3 mg) to stimulate uterine PGF2 alpha production. Plasma concentrations of progesterone (P4) and 15-keto-13,14-dihydro-PGF2 alpha (PGFM) were determined by RIA. The PGFM responses following E2 challenge were decreased (p less than 0.01) for cows receiving CSP versus serum proteins into the uterine lumen. Individual PGFM, P4 and cycle length responses are discussed. Data suggest that proteins secreted by the bovine conceptus suppress uterine PGF2 alpha production during pregnancy recognition in the cow. 相似文献
170.