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991.
Replication of bacteriophage lambda DNA. Examination of variants containing double origins and observation of a bias in directionality 总被引:1,自引:0,他引:1
Maria Schnös Katherine J. Denniston Frederick R. Blattner Ross B. Inman 《Journal of molecular biology》1982,159(3):441-455
Bacteriophage λ variants have been constructed that possess two λ ori sites. Replicative intermediates resulting from infection with these phages have been investigated. We find that initiation of replication from the ori site on an EcoRI fragment (containing all the DNA sequences from within the red gene to the middle of gene O) cloned in the inverted orientation is predominantly bidirectional but occurs at a decreased frequency. Double initiations were observed at low frequency. However, a second cloned ori fragment (carrying two large deletions and a small insertion) cloned in the normal orientation demonstrated insignificant levels of replication from the cloned site unless the normal ori had already initiated.A bias in directionality of λ replication has been observed. Molecules that replicate unidirectionally propagate to the right more often than to the left. If the cloned ori-containing EcoRI fragment is inserted with reversed polarity, then the bias is towards the left. Bidirectional λ replicative intermediates also appear to show a similar bias but this is superimposed on a large, apparently random, effect that results in asymmetric growing-point propagation. 相似文献
992.
Summary Transepithelial current fluctuations were recorded inNecturus gallbladder, clamped at negative as well as positive potentials up to 64 mV. With NaCl-Ringer's (+10mm TAP) on both sides a mucosa-negative potential enhanced the relaxation noise component, present at zero potential, and produced peaking in the power spectrum at potentials above –36mV. Concomitantly at these potentials an inductive as well as a capacitive low-frequency feature appeared in the impedance locus. Clamping at positive potentials of 18 mV suppressed the relaxation noise component. At potentials above 51mV the spectral values increased predominantly at low frequencies. In this case the power spectrum showed only a 1/f
noise component. The experiments confirm the previous finding that a K+ efflux through fluctuating apical K+ channels exists under normal conditions. With serosal KCl-Ringer's the initial Lorentzian component was enhanced at negative but suppressed at positive potentials. The increase at negative potentials was less pronounced than in experiments with NaCl-Ringer's on both sides, indicating saturation of the fluctuating K+ current component. With mucosal KCl-Ringer's a negative potential depressed the initial relaxation noise component, whereas it was enhanced at +18 mV clamp potential. In the latter case an additional Lorentzian component became apparent at higher frequencies. At potentials of 36 mV and above the low-frequency Lorentzian disappeared whereas the corner frequency of the high-frequency component increased. The latter experiments demonstrate that the relaxation noise component inNecturus gallbladder consists of two superimposed Lorentzians. As the relaxation times of these two components behave differently under an electrical field, there may exist two different types of K+ channels. It is demonstrated that peaking in the plateau of power spectra can be explained by frequency-dependent attenuation effects, caused by a polarization impedance. 相似文献
993.
W.F. Anderson M.G. Grütter S.J. Remington L.H. Weaver B.W. Matthews 《Journal of molecular biology》1981,147(4):523-543
Phage lysozyme has catalytic activity similar to that of hen egg white lysozyme, but the amino acid sequences of the two enzymes are completely different.The binding to phage lysozyme of several saccharides including N-acetylglucosamine (GlcNAc), N-acetylmuramic acid (MurNAc) and (GlcNAc)3 have been determined crystallographically and shown to occupy the pronounced active site cleft. GlcNAc binds at a single location analogous to the C site of hen egg white lysozyme. MurNAc binds at the same site. (GlcNAc)3 clearly occupies sites B and C, but the binding in site A is ill-defined.Model building suggests that, with the enzyme in the conformation seen in the crystal structure, a saccharide in the normal chair configuration cannot be placed in site D without incurring unacceptable steric interference between sugar and protein. However, as with hen egg white lysozyme, the bad contacts can be avoided by assuming the saccharide to be in the sofa conformation. Also Asp20 in T4 lysozyme is located 3 Å from carbon C(1) of saccharide D, and is in a position to stabilize the developing positive charge on a carbonium ion intermediate. Prior genetic evidence had indicated that Asp20 is critically important for catalysis. This suggests that in phage lysozyme catalysis is promoted by a combination of steric and electronic effects, acting in concert, The enzyme shape favors the binding in site D of a saccharide with the geometry of the transition state, while Asp20 stabilizes the positive charge on the oxocarbonium ion of this intermediate. Tn phage lysozyme, the identity of the proton donor is uncertain. In contrast to hen egg white lysozyme, where Glu35 is 3 Å from the glycosidic DOE bond, and is in a non-polar environment, phage lysozyme has an ion pair, Glull … Arg145, 5 Å away from the glycosidic oxygen. Possibly Glull undergoes a conformational adjustment in the presence of bound substrate, and acts as the proton donor. Alternatively, the proton might come from a bound water molecule. 相似文献
994.
Laura de Magistris Gianfranco Delle Fave Anna Kohn Thue W. Schwartz 《Life sciences》1981,28(23):2617-2621
Bombesin, besides many other actions on the mammalian gastroentero-pancreatic tract, strongly stimulates the release of pancreatic-polypeptide (PP) in dogs. In 8 healthy human volunteers (5 males, 3 females), the PP response during bombesin infusion was low (25.7 ± 6.3 peak vs. 5.0 ± 2.0 basal pmol/1) compared to the effect of a protein meal (144.1 ± 13.4 pmol/1) or to the gastrin response to the same dose of the amphibian polypeptide (140.0 ± 23.6 pmol/1 eq SHG 17 I). The response pattern of PP and gastrin was different as PP concentrations peaked 10 min after cessation of bombesin infusion (32.0 ± 4.9 pmol/1) when gastrin concentrations already were down to one third of the maximal response. Atropine inhibited the PP response to bombesin but did not abolish it completely. It is concluded that in man, the total effect of bombesin on PP secretion is minor compared both to the effect of the peptide on gastrin secretion in man and to the effect of bombesin in dogs. It is suggested that bombesin might have a dual, inhibitory-stimulatory, effect on PP secretion in man. 相似文献
995.
This paper presents a generalization of the classical compartmental models using the concept of "pipes" between the compartments and taking into account the time needed for the material to pass through these pipes. A general linear delay differential system describing such models is derived and some properties of its solutions-such as nonnegativity, boundedness, stability-are established. For stationary and asymptotically stationary systems the asymptotic solution is obtained, too. The theory presented generalizes several earlier results concerning more special equations. 相似文献
996.
The amounts of AMP, ADP and ATP were determined after sulfite incubation in photo-organotrophically cultivated cells of the green alga Trebouxia. D 相似文献
997.
Summary Ectomycorrhizae were found in root samples of the treesAfzelia quanzensis Welw. andBrachystegia spiciformis Benth. (Caesalpiniaceae), collected in the coastal miombo type woodland 50 km west of Dar-es-Salaam, Tanzania. Root nodules
with a structure resembling that of nitrogen-fixing root nodules of other leguminous plants were observed in theA. quanzensis material. The climate of the locality is rather dry, and strongly seasonal. In the tropics, ectomycorrhizae have previously
been found only in humid or rain forest climate zones. 相似文献
998.
Effect of abscisic acid on the transport of assimilates in barley 总被引:15,自引:0,他引:15
The effect of abscisic acid (ABA) on assimilate transport in barley was investigated in two parallel experiments. First, the effect upon [14C]sucrose transport from the flag leaf to the ear of a single ABA application made at different stages of growth of the fruits was investigated; the effect was measured 24 h after treatment. Second, the effect of a single application of ABA made at the same stages of growth as above on grain weight of the mature plant was investigated. In both types of experiments ABA was applied once to the ear of different plants as an aqueous solution (10-3–10-5 M), one to five weeks after anthesis. [14C] sucrose was applied by means of agar blocks. Parallel to these experiments, the endogenous content of ABA was investigated in the developing grains. When ears were treated with ABA two or four weeks after anthesis, an increase of up to 70% in the 14C-transport from the flag leaf to the ear was observed within a 24-h period after treatment (short duration experiments). At these growth stages the endogenous concentrations of ABA were low. In sharp contrast, ABA, especially in a concentration of 10-3 M, decreased 14C-import from the flag leaf when applied three weeks after anthesis. At this stage the endogenous ABA content had reached its maximum. Long duration experiments with a single application of ABA to the car two weeks after anthesis resulted in a marked increase of weight per thousand kernels. ABA applications made earlier or later than two weeks after anthesis either reduced the grain weight or had no effect. It is concluded that ABA is involved in the regulation of assimilate transport from the leaves to the grains, possibly by influencing the unloading of sieve tubes in the ears. Promotion or inhibition of assimilate import by exogenously applied ABA may depend on the developmental stage of the grains and on the endogenous ABA level.Abbreviations ABA
abscisic acid
- TKW
weight per thousand kernels 相似文献
999.
The cofactor of enzymatic, 1-aminocyclopropane-1-carboxylic acid dependent ethylene formation was concentrated on cation exchange columns. When chelators of cations were added to the homogenates, cofactor activity was lost. Cofactor fractions were partly resistant to oxidation at 600° C. Mn2+ substituted for the cofactor in ethylene formation from 1-aminocyclopropane-1-carboxylic acid by a protein fraction isolated from etiolated pea shoots. In addition, Mn2+ enhanced the stimulatory effect of the concentrated cofactor. The elution volume for the cofactor on a Sephadex G-25 column was lower than that of MnCl2. In paper electrophoresis the cofactor migrated to the cathode at pH 10.8 and 2.2. The RF of cofactor on cellulose plates developed in butanol: acetic acid: H2O was 0.4. After cellulose chromatography, cofactor activity had to be reconstituted by the addition of MnCl2. Chelators, anti-oxidants, and catalase were inhibitors of Mn2+-cofactor-dependent ethylene formation. The protein necessary for 1-aminocyclopropane-1-carboxylic acid dependent ethylene formation in vitro was seperated from 95–98% of the total protein in homogenates by DE-52 cellulose chromatography and (NH4)2SO4-fractionation.Abbreviations ACC
1-aminocyclopropane-1-carboxylic acid
- EDTA
ethylenediaminetetraacetic acid
- DDTC
diethyldithiocarbamate 相似文献
1000.
Calcified Mandl's corpuscles present in the internal layer (or fibrillary plate) of the teleost fish scale were studied by transmission and scanning electron microscopy for a better understanding of this special type of mineralization process. The corpuscles show a great variability in their structure, form and surface features depending on the arrangement of the collagen fibrils in the internal layer of the different fish species studied, on the localization of the corpuscles in the scale and on the technical treatment to which the scale is subjected. 相似文献