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61.
Summary Rare mutations that alter the substrate specificity of proline permease cluster in discrete regions of theputP gene, suggesting that they may replace amino acids at the active site of the enzyme. IfputP substrate specificity mutations directly alter the active site of proline permease, the mutants should show specific defects in the kinetics of proline transport. In order to test this prediction, we examined the kinetics of threeputP substrate specificity mutants. One class of mutation increases theK
m
over 120-fold but only decreases theV
max fourfold. SuchK
m
mutants may be specifically defective in substrate recognition, thus identifying an amino acid critical for substrate binding. Another class of mutation decreases theV
max 80-fold without changing theK
m
.V
max mutants appear to alter the rate of substrate translocation without affecting the substrate binding site. The last class of mutation alters both theK
m
andV
max of proline transport. These results indicate that substrate specificity mutations alter amino acids critical for Na+/proline symport. 相似文献
62.
Carbon: terrestrial C4 plants 总被引:1,自引:1,他引:0
The carbon isotope composition of terrestrial C4 plants depends on the primary carboxylation of phosphoenolpyruvate (PEP) and on the diffusion of CO2 to the carboxylation sites, but is also influenced by the final carboxylation of ribulose-1,5-bisphosphate (RuBP). Several models have been used for reproducing this complex situation. In the present review, a particular model is applied as a means to interpret the effects of environmental and genetically determined factors on carbon isotope discrimination during C4 photosynthesis. As a new feature, the model considers four types of limitation of the overall CO2 assimilation rate. Both carboxylation reactions are assumed to be limited by either maximum enzyme activity or maximum substrate regeneration rate. The model is applied to experimental data on the effects of CO2, irradiance and water stress on short-term discrimination by leaves of several C4 species measured simultaneously with CO2 gas exchange characteristics. In particular, different patterns of the influence of low irradiances on carbon isotope discrimination are interpreted as due to variations in that irradiance at which a transition from limitation by PEP regeneration rate and RuBP carboxylase activity to limitation by the regeneration rates of both substrates occurs. After discussing literature data on the effects of environmental conditions on carbon isotope discrimination by C4 plants seasonal and developmental changes in carbon isotope composition, studies on the systematic and geographic distribution of C4 plants, evolutionary and genetical aspects, and some ecological implications are reviewed. 相似文献
63.
Transformed 3T3 cells have reduced levels and altered subcellular distribution of the major PKC substrate protein marcks 总被引:1,自引:0,他引:1
The MARCKS (myristylated alanine-rich C-kinase substrate) protein is an abundant calmodulin-binding protein that is a major and specific endogenous substrate of protein kinase C (PKC). Stimulation of cells with phorbol esters or other activators of PKC has been shown previously to result in rapid phosphorylation of MARCKS proteins and redistribution of these myristylated C-kinase substrates from membrane to cytosol. Here we show that NIH3T3 murine fibroblasts transformed by p21-HA-C-RAS or pp60-V-SRC oncoproteins have markedly reduced levels of p68-MARCKS and that most of the remaining MARCKS protein is found in the cytosol. 3T3 cells containing a nontransforming oncoprotein p26-BCL2, in contrast, exhibited normal levels and distribution of p68-MARCKS. When taken together with recent evidence that MARCKS proteins are involved in regulating organization of the membrane cytoskeleton, our findings suggest that oncoprotein-mediated alterations in MARCKS protein levels and subcellular distribution may contribute to the development or maintenance of the transformed phenotpe. 相似文献
64.
L. P. Ruse 《Aquatic Ecology》1992,26(2-4):411-417
Chironomid pupal skins were collected during one year from three sites along a chalk stream in southern England. The sites had similar chemistry and discharge but differed in their current and temperature regimes. Substrate composition upstream of each collection point was surveyed during spring, summer and autumn. Proportions of macrophytes and sediments were compared with proportions of chironomid species and trophic groups. Seasonal changes in substrate and pupal skin collections were correlated by classification and direct ordination techniques. The distribution of chironomid species were indicated as being significantly related to the recorded substrate data. 相似文献
65.
Macroinvertebrate density, biomass and drift were studied from moss-covered and moss-free channels in the South Fork Salmon River, Idaho. Insect densities were compared for 10 different substrate types and locations involving moss (Fontinalis neo-mexicana), sand, pebbles and cobbles. An ANOVA test demonstrated that insect densities varied significantly with substrate type (P < 0.05), and that total insect density in moss clumps differed significantly from densities in mineral substrates. Insect densities were 4–18 times greater in moss clumps than in mineral substrates under and adjacent to moss; sands under moss supported the lowest densities. During most tests, densities in pebble and cobble substrates adjacent to moss clumps were not significantly different from those found in similar substrates in the moss-free channel. The 20% moss-covered channel had 1.6 to 7.2 greater insect density and 1.4 to 6.1 greater biomass than did the moss-free channel for the tests conducted. Generally, midges (Chironomidae) made up over 50% of the insect community; annelids were the principal non-insect invertebrates.In spite of greater insect density and biomass in a moss-covered than in the moss-free channel, we did not demonstrate universally increased drift of the immature stages from the moss-covered channel, at least during daylight hours. As a consequence, we infer that salmonid fishes, feeding primarily on drifting insects during the daytime, may not derive increased caloric benefit from moss habitats until the insects emerge as adults. 相似文献
66.
The degradation of phenol by Rhodococcus sp. P1 was studied in continuous culture systems. The organism could be adapted by slowly increasing concentration, step by step, up to 30.0 g · 1-1 phenol in the influent. The degradation rate reached values of about 0.3 g · g dry mass-1 ·h-1. Large step increases in phenol concentration and addition of further substrates (e.g., catechol) were tolerated up to a certain concentration. With increasing dilution rate and increasing inlet phenol concentration the stability of the system decreased. 相似文献
67.
Horse liver contains previously unrecognized isozymes of alcohol dehydrogenase. In contrast to the molecular forms identified up to now, under the conditions employed these variants migrate toward the anode on starch gel electrophoresis and were separated from the cathodic isozymes by DEAE-cellulose chromatography. They were then purified on agarose-hexane-AMP. Their physicochemical and compositional characteristics are similar to those x alcohol dehydrogenases from human liver. Like these and similar ones from simian liver, they retain most of their activity in the presence of10
mm 4-methylpyrazole, oxidize short-chain primary alcohols very poorly, and appear to prefer longer chain primary alcohols and -hydroxy fatty acids as substrates. 相似文献
68.
The growth of Hansenula polymorpha and Kloeckera sp. 2201 with a mixture of glucose and methanol (38.8%/61.2%, w/w) and the regulation of the methanol dissimilating enzymes alcohol oxidase, catalase, formaldehyde dehydrogenase and formate dehydrogenase were studied in chemostat culture, as a function of the dilution rate. Both organisms utilized and assimilated glucose and methanol simultaneously up to dilution rates of 0.30 h-1 (H. polymorpha) and 0.26h-1, respectively (Kloeckera sp. 2201) which significantly exceeded max found for the two yeasts with methanol as the only source of carbon. At higher dilution rates methanol utilisation ceased and only glucose was assimilated. Over the whole range of mixed-substrate growth both carbon sources were assimilated with the same efficiency as during growth with glucose or methanol alone.In cultures of H. polymorpha, however, the growth yield for glucose was lowered by the unmetabolized methanol at high dilution rates. During growth on both carbon sources the repression of the synthesis of all catabolic methanol enzymes which is normally caused by glucose was overcome by the inductive effect of the simultaneously fed methanol. In both organisms the synthesis of alcohol oxidase was found to be regulated differently as compared to catalase, formaldehyde and formate dehydrogenase. Whereas increasing repression of the synthesis of alcohol oxidase was found with increasing dilution rates as indicated by gradually decreasing specific activities of this enzyme in cell-free extracts, the specific activities of this enzyme in cell-free extracts, the specific activities of catalase and the dehydrogenases increased with increasing growth rates until repression started. The results indicate similar patterns of the regulation of the synthesis of methanol dissimilating enzymes in different methylotrophic yeasts.Abbreviations and Terms C1
Methanol
- C6
glucose; D dilution rate (h-1)
-
D
c
critical dilution rate (h-1)
-
q
s
specific, rate of substrate consumption (g substrate [g cell dry weight]-1 h-1)
-
q
CO2 and q
O2
are the specific rates of carbon dioxide release and oxygen consumption (mmol [g cell dry weight]-1 h-1)
- RQ
respiration quotient (q
CO2
q
O2
1
)
-
s
0(C1) and s
0(C6)
are the concentrations of methanol and glucose in the inflowing medium (g l-1)
-
s
residual substrate concentration in the culture liquid (g l-1)
- Sp. A.
enzyme specific activity
- x
cell dry weight concentration (gl-1)
-
Y
X/C6
growth yield on glucose (g cell dry weight [g substrate]-1 相似文献
69.
Koichi Hirata Tadashi Oku Aaron E. Freeman 《In vitro cellular & developmental biology. Plant》1982,18(9):789-799
Summary Twenty to twenty-two days postcoitum mouse fetal pancreas organ bits were cultured on the dermal surface of irradiated pigskin
as a substrate. The medium used for long term culture consisted of Eagle’s Minimum Essential Medium with the addition of 10%
bovine serum, 0.02 U/ml insulin, 0.025 μg/ml glucagon, 3.63 μg/ml hydrocortisone, 100 μg/ml soybean trypsin inhibitor or 10−8
M atropine. When the medium lacked trypsin inhibitor or atropine but contained the three hormones, the pigskin support began
to be destroyed after 2 to 4 wk in culture. Thereafter, the cultured cells could not grow and survive on the digested pigskin.
When 10−6
M atropine was added to the medium, amylase secretion from cultured cells and destruction of pigskin were inhibited completely
but pancreas cells could not grow or survive. In contrast, 100 μg/ml soybean trypsin inhibitor or 10−8
M atropine permitted cell growth, permitted amylase secretion from the cultured acinar cells, and prevented the destruction
of pigskin. Under these conditions pancreas cells migrated or grew or both from the organ bits onto the surface of the pigskin
dermis and organoid aggregations formed. Hydrocortisone was needed to permit growth for more than 2 wk. Glucagon and insulin
had additive effects. Light and electron microscopic observations indicated the culture of at least five kinds of cells, i.e.,
duct, acinar, centroacinar, endocrine, and mesenchymal. The majority of cultured cells were duct cells and acinar cells. There
were few mesenchymal cells. Mouse pancreas cells were cultured for at least 12 wk by this method.
This investigation was supported by PHS Grant CA 30220 awarded by the National Cancer Institute, DHHS, Grant 1203M awarded
by the Council for Tobacco Research, Inc., and Grant RD-65 (for equipment) awarded by the American Cancer Society. Nude mice
were provided by Dr. Wendall M. Farrow of Life Sciences, Inc., Resource Laboratory N01, CP6-1005 of the National Cancer Institute. 相似文献
70.
D. Dudley Williams 《Hydrobiologia》1981,83(2):257-260
The relative efficiencies of two sizes of a standpipe corer were evaluated. The size composition of the gravel sampled by the corer was very similar to that (below the opening size of the core chamber) of the streambed. The small corer (25 cm3 sample size) produced a mean overestimate of total numbers of only 19% even in highly heterogeneous gravels. Most of the taxa commonly in the substrates sampled did not escape from the corer. A few rare taxa were consistently over- or under-estimated and possible reasons for this are discussed. 相似文献