全文获取类型
收费全文 | 14209篇 |
免费 | 1412篇 |
国内免费 | 9篇 |
专业分类
15630篇 |
出版年
2022年 | 109篇 |
2021年 | 199篇 |
2020年 | 121篇 |
2019年 | 145篇 |
2018年 | 185篇 |
2017年 | 173篇 |
2016年 | 292篇 |
2015年 | 570篇 |
2014年 | 597篇 |
2013年 | 739篇 |
2012年 | 898篇 |
2011年 | 936篇 |
2010年 | 651篇 |
2009年 | 534篇 |
2008年 | 775篇 |
2007年 | 833篇 |
2006年 | 808篇 |
2005年 | 785篇 |
2004年 | 728篇 |
2003年 | 706篇 |
2002年 | 753篇 |
2001年 | 148篇 |
2000年 | 111篇 |
1999年 | 174篇 |
1998年 | 207篇 |
1997年 | 160篇 |
1996年 | 145篇 |
1995年 | 133篇 |
1994年 | 124篇 |
1993年 | 137篇 |
1992年 | 112篇 |
1991年 | 112篇 |
1990年 | 126篇 |
1989年 | 116篇 |
1988年 | 109篇 |
1987年 | 117篇 |
1986年 | 94篇 |
1985年 | 104篇 |
1984年 | 119篇 |
1983年 | 124篇 |
1982年 | 147篇 |
1981年 | 124篇 |
1980年 | 101篇 |
1979年 | 71篇 |
1978年 | 86篇 |
1977年 | 75篇 |
1976年 | 68篇 |
1975年 | 61篇 |
1974年 | 69篇 |
1973年 | 55篇 |
排序方式: 共有10000条查询结果,搜索用时 15 毫秒
31.
Characterisation of a new monoclonal antibody (mAb), designated LAS 41, directed against 124-kilodalton (kDa) etiolated-oat (Avena sativa L.) phytochrome, indicates that it recognises an epitope unique to the red-light-absorbing form, Pr. In a solid-phase enzyme-linked immunosorbent assay (ELISA), LAS 41 exhibits a seven- to eight-fold higher affinity for Pr than for the far-red-light-absorbing form of phytochrome, Pfr. In addition, in immunoprecipitation assays LAS 41 effectively precipitates 100% of phytochrome presented as Pr but only precipitates a maximum of 24.5% of phytochrome presented as Pfr. These values are indicative of binding exclusively to Pr. Peptide-mapping studies show that LAS 41 recognises and epitope located within a region 6–10 kDa from the aminoterminus of the phytochrome molecule. Since binding of LAS 41 to Pr induces alterations in the spectral properties of Pr, this indicates that at least part of the 4 kDa domain to which the antibody binds is essential for protein-chromophore interaction. Subsequent photoconversion of LAS 41-Pr complexes produces native Pfr spectra, with concomitant production of free antibody and antigen, as shown by a modified ELISA. The specificity of LAS 41 for Pr has facilitated the purification of Pfr which is free of contaminating Pr. This has enabled direct determination of the mole fraction of Pfr established by red light to be 0.874.Abbreviations ELISA
enzyme-linked immunsorbent assay
- kDa
kilodalton
- mAb
monoclonal antibody
- Pfr
far-red-absorbing form of phytochrome
- Pr
red-absorbing form of phytochrome
- SDS-PAGE
sodium dodecyl sulphate polyacrylamide gel electrophoresis
- (A)
difference in absorbance (A
665
Pr
–A
730
Pr
)-(A
665
Pfr
–A
730
Pfr
)
- Ar/Afr
spectral change ratio (SCR)
- max
mole fraction of Pfr following saturating red light 相似文献
32.
We found that extensive metabolic switching can be triggered in aromatization. Up to 20–35% of (19-3H3)-labeled substrate is diverted to non-estrogenic product, namely 1β- and 2β-hydroxy androgens, negating usefulness for [3H]water assays. The strikingly substrate-specific metabolite patterns observed reflect positioning at the active site, and the large kinetic isotope effects involved. This switching is unusual in that it involves: 1) tritium labeling, 2) a multistep enzymic process, and 3) the potential unraveling of an enzymic mechanism. 相似文献
33.
Stanley A. Vinores Mary M. Herman Lucien J. Rubinstein 《The Histochemical journal》1987,19(8):439-448
Summary Neuron-specific () enolase, a glycolytic enzyme used as a relatively specific marker for normal neurons and neuroendocrine cells, has recently been found in a variety of neoplastic cells and in reactive astrocytes. Its localization was investigated by immunohisto- and electron-immunocyto-chemistry, in the proliferating supportive Schwann cells of a peripheral ganglioneuroblastoma and in the neoplastic Schwann cells of four acoustic tumours. By light microscopy, the neoplastic Schwann cells showed moderate uneven diffuse immunopositivity for enolase. By electron-immunocy-tochemistry, both types of Schwann cells demonstrated immunopositivity discretely limited to their cell surface membranes. The neoplastic ganglion cells and axons of the ganglioneuroblastoma and the normal neurons and axons included in the schwannomas were, as expected, intensely immunopositive. The visualization of enolase on the cell surface membranes of both neoplastic and non-neoplastic proliferating Schwann cells suggests that increased glycolytic activity may occur on the surface of these proliferating cells irrespective of the nature of the proliferation. 相似文献
34.
Beth J. Hoffman Ursula Scheffel† John R. Lever† Michael D. Karpa Paul R. Hartig† 《Journal of neurochemistry》1987,48(1):115-124
Methylation of 2-125I-lysergic acid diethylamide (125I-LSD) at the N1 position produces a new derivative, N1-methyl-2-125I-lysergic acid diethylamide (125I-MIL), with improved selectivity and higher affinity for serotonin 5-HT2 receptors. In rat frontal cortex homogenates, specific binding of 125I-MIL represents 80-90% of total binding, and the apparent dissociation constant (KD) for serotonin 5-HT2 receptors is 0.14 nM (using 2 mg of tissue/ml). 125I-MIL also displays a high affinity for serotonin 5-HT1C receptors, with an apparent dissociation constant of 0.41 nM at this site. 125I-MIL exhibits at least 60-fold higher affinity for serotonin 5-HT2 receptors than for other classes of neurotransmitter receptors, with the dopamine D2 receptor as its most potent secondary binding site. Studies of the association and dissociation kinetics of 125I-MIL reveal a strong temperature dependence, with very slow association and dissociation rates at 0 degree C. Autoradiographic experiments confirm the improved specificity of 125I-MIL. Selective labeling of serotonin receptors was observed in all brain areas examined. In vivo binding studies in mice indicate that 125I-MIL is the best serotonin receptor label yet described, with the highest frontal cortex to cerebellum ratio of any serotonergic radioligand. 125I-MIL is a promising ligand for both in vitro and in vivo labeling of serotonin receptors in the mammalian brain. 相似文献
35.
Carbohydrate components (simple sugars and polysaccharides) of cell walls of pearl millet (Pennisetum americanum L., cv. Gahi) were studied as potential substrates for the root-associated diazotroph Azospirillum brasiliense Sp. 7. Simple sugars were utilized, but no evidence was obtained to support the suggestion that the polysaccharide components tested might serve as substrates for growth following hydrolysis by the associated azospirilla. 相似文献
36.
A Mary K E Achyuthan C S Greenberg 《Biochemical and biophysical research communications》1987,147(2):608-614
The binding sites in fibrinogen for Factor XIII were localized using an immunoblotting technique. Platelet Factor XIII bound to fibrinogen and to plasmin degradation products of fibrin(ogen) including Fragments: X, D1-D3, and D-dimer, but did not bind to Fragment E. Binding of Platelet Factor XIII was independent of calcium ions but could be inhibited by the presence of 0.5 M NaCl. Binding could also be inhibited by preincubating Factor XIII with a 100-fold molar excess of fibrinogen but not by 100-fold molar excess of Fragment E. Binding of Factor XIII to fibrinogen was specific, since several other proteins tested (ovalbumin, bovine serum albumin, alpha 2-macroglobulin, beta-galactosidase, fructose kinase, lactic dehydrogenase, triose phosphate isomerase, fumarase and pyruvate kinase) did not bind Factor XIII. Furthermore, binding was not observed either when Factor XIII was left out or when antiFactor XIII antiserum was substituted with nonimmune serum. When fibrinogen was reduced prior to electrophoresis, Factor XIII bound to the A alpha and B beta chains of fibrinogen and des A,B fibrinogen, the B beta-chain of Fragment X, but not the gamma-chains. Localization of the Factor XIII binding sites to the carboxy terminal segments of the A alpha and B beta chains in the Fragment D-domain of fibrinogen could have important physiological consequences. 相似文献
37.
This paper contains new experimental data on the growth dynamicsof a lobate coastal ctenophore, Mnenmiopsis mccradyi, whichadd significantly to our understanding of the nutritional ecologyof ctenophores and their role as opportunistic predators. Theseexperimental observations were necessary to refine the dynamiccarbon budget presented as a simulation model in another report.The ratio of carbon biomass to dry weight may vary several-folddepending on the nutritional state and size from >12% inwell-fed larvae to <1% in starved adults. Feeding effort(clearance rate) is higher for previously starved animals, fallingsharply within a few hours after re-exposure to food. Directvisual observations of feeding activity of animals confirmedthis. Assimilation efficiency can be high (72%) in these animalsbut if they continue to feed at high food concentrations, incomingfood displaces material which is only partially digested andassimilation efficiency decreases substantially. Except at verylow food concentrations, growth efficiency ranges between 20and 45%. Mnemiopsis, begins to produce eggs at a size much lessthan its maximum. Egg production is very sensitive to food supply,and somatic growth is favored over egg production at low fooddensities. Even though several thousand eggs may be producedover a few days, they represent <2% day1 of the carbonbiomass of the ctenophore and several-fold less than respiratorycarbon. 相似文献
38.
We have microinjected DNA containing the inducible mouse metallothionein-I (MT-I) promoter, coupled to the structural gene for Escherichia coli β-galactosidase (lacZ), into the pronuclei of one-cell mouse embryos. A qualitative histochemical assay, with 5-bromo-4-chloro-3-indolylβ-
-galactopyranoside (X-Gal) as a substrate, was used to detect expression of lacZ at several preimplantation stages. We observed staining indicative of exogenous β-galactosidase activity in 5–17% of DNA-injected embryos assayed at preimplantation stages after 16–24 h treatment with ZnSO4. Thus, lacZ can be used as an indicator gene for promoter function during early mouse embryogenesis, and the incorporation of the MT-I promoter into fusion genes can be a useful means of controlling the expression of exogenous genes in preimplantation mouse embryos. 相似文献
39.
Mary E. Lidstrom 《Antonie van Leeuwenhoek》1989,55(1):7-14
Summary By using molecular probe techniques in combination with activity and expression measurements, it is possible to estimate bacterial
populations in nature. This information can be expooited to study a number of important environmental problems. For instance,
it will be possible to study ecosystem perturbation and microbial competition, by altering an ecosystem or a laboratory model
of an ecosystem, and assessing corresponding changes in key activities and populations. In addition, regulation of activities
in the laboratory can be compared to the response of activities and populations in situ, to develop an understanding of the
key parameters that control these processes in nature. These types of approaches are important steps for determining the role
of microorganisms in geochemical cycling, in both specific habitats and on a global basis. 相似文献
40.
Biotransformations of aromatic aldehydes by acetogenic bacteria 总被引:2,自引:0,他引:2
Mary F. Luxa Elizabeth Keitha Tsungda Hsua Harold L. Drakea 《FEMS microbiology letters》1990,67(1-2):73-78
Vanillin was subject to O demethylation and supported growth of Clostridium formicoaceticum and Clostridium thermoaceticum. Vanillin was also stimulatory to the CO-dependent growth of Peptostreptococcus productus. The aldehyde substituent of vanillin was metabolized by routes which were dependent upon both the acetogen and a co-metabolizable substrate (e.g. carbon monoxide [CO]). C. formicoaceticum and C. thermoaceticum oxidized the aldehyde group of vanillin to the carboxyl level, while P. productus reduced the aldehyde group of vanillin to the alcohol level. In contrast, during CO-dependent growth, C. thermoaceticum reduced 4-hydroxybenzaldehyde to 4-hydroxybenzyl alcohol while P. productus both reduced and oxidized 4-hydroxybenzaldehyde to 4-hydroxybenzyl alcohol and 4-hydroxybenzoate, respectively. These metabolic potentials indicate aromatic aldehydes may affect the flow of reductant during acetogenesis. 相似文献