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61.
A A da Gama 《Journal of theoretical biology》1990,142(2):251-260
A model for the through-bond electronic interaction between electron donor and acceptor in proteins is developed. We use a one-electron Hamiltonian, write the Dyson's equation in site representation and solve it by using a Green's function formalism with some renormalization ideas. An expression for Tab which describes the exponential decay with distance bond per bond is obtained. Covalent, non-covalent and convergent pathways are considered and no periodic approximation is needed. 相似文献
62.
M N Simon T Winckler R Mutzel M Véron J C da Costa Maia 《Biochemical and biophysical research communications》1992,184(3):1142-1151
Extracts from Dictyostelium discoideum contain type 2A and 2C serine/threonine-specific protein phosphatases with properties very similar to those from mammals according to their sensitivity to okadaic acid and to their dependence for divalent cations. In contrast, no type 1 protein phosphatase is found at any time of development, neither in the cytosolic nor in the particulate fraction, using glycogen phosphorylase a, casein, histone or the non-proteinous 4-Methylumbelliferyl phosphate as substrates. Both type 2A and 2C protein phosphatase activities remain constant throughout the development cycle. 相似文献
63.
We compared the transient activity of three cereal gene-derived promoter-gus fusions and the efficiency of selection mediated by three different selectable genes in a polyethylene glycol transformation system with haploid cell suspension protoplasts of rice. The maize ubiquitin promoter was found to be the most active in transformed protoplasts, and selection on ammonium glufosinate mediated by the bar gene was the most efficient for producing resistant calluses. Cotransformation of protoplasts with two separate plasmids carrying the gus and the bar genes, at either a 21 or 11 ratio, led to 0.8 × 10–5 and 1.6 × 10–5 resistant callus recovery frequencies and 59.7 and 37.9 cotransformation efficiencies respectively. No escapes were detected in dot blot analyses of 100 resistant calluses with a probe consisting of the bar coding region. Cotransformation efficiency, based on resistance to basta and -glucuronidase staining of the leaf tissue of 115 regenerated plants, was 47%. Resistance tests and Southern analysis of seed progenies of three diploid transgenic plants demonstrated homozygous integration of multiple copies of the transgene at one locus at least in the first plant, heterozygous integration at one locus in the second plant and heterozygous integration at two loci in the third plant.Abbreviations PEG
polyethylene glycol
- T0
regenerated transgenic plant
- GUS
-glucuronidase
- CaMV
cauliflower mosaic virus
- ARE
anaerobic responsive element
- OCS
octopine synthase
- T1
first generation progeny of transgenic plants 相似文献
64.
We describe the application of lamin immunocytochemistry (ICC) and single- or double-target fluorescence in situ hybridization (FISH) on 4 microm thick frozen tissue sections as a method to facilitate scoring of aberrant chromosome copy numbers in colonic tumors. Analysis of FISH signals in colon tissue sections is often hampered by overlap and truncation of epithelial nuclei, due to the density of the epithelial cells. Furthermore, on the basis of nuclear staining it is often difficult to determine whether or not nuclei are overlapping, or adjoining. Therefore, reliable evaluation of (F)ISH signals to screen for genomic changes was until now mainly restricted to isolated nuclei obtained from relatively thick tissue sections. In this study the applicability of lamin ICC, to stain the nuclear periphery and to distinguish individual nuclei, combined with the FISH procedure is explored to solve this problem for colon epithelium. For ICC we applied the alkaline phosphatase (APase)-Fast Red detection method, since the fluorescent precipitate of this reaction resists extensive proteolytic digestion as needed for efficient FISH on tissue sections. Chromosome copy numbers could easily be determined in 4 microm thick frozen tissue sections by combining lamin ICC and FISH. The ratio of the copy numbers of the chromosomes 7 and 17 could be determined in frozen tissue sections after combined lamin ICC and double-target FISH. It is concluded that the combination of lamin ICC and FISH improves chromosome copy number analysis and can be used to investigate genomic changes in different tumor compartments in thin frozen tissue sections. 相似文献
65.
The majora2–6 sialoglycoproteins in detergent-extracts of Kurloff cells were purified by anion-exchange andSambucus nigra agglutinin-affinity chromatographies. The similar ultrastructural localisations of (1)S. nigra agglutinin-gold conjugates and (2) acid phosphatase activities on the Kurloff body and particularly on its myelin figures indicated that the majora2-6 sialoglycoproteins of the Kurloff cell had acid phosphatase activity. Two-dimensional electrophoresis showed that these tartrate-sensitive phosphatases corresponded to 2 acidic (pI 3.4–3.7) polypeptides of 36 and 34 kDa. Hydrolysis with peptide-N-glycosidases F gave a 33 kDa apoprotein rich in alanine, glutamic acid, tyrosine and lysin. A lectin-affinity study demonstrated that they contained hybrid type bisected and fucosylatedN-linked oligosaccharides. Cytotoxic properties were previously attributed to Kurloff cells and other studies suggested that not only acid phosphatases but alsoa2-6-linked sialic acid residues themselves may participate in natural killer activity. 相似文献
66.
Markers for selection of the rice Xa21 disease resistance gene 总被引:8,自引:0,他引:8
C. E. Williams B. Wang T. E. Holsten J. Scambray F. de Assis Goes da Silva P. C. Ronald 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1996,93(7):1119-1122
Six molecular markers were mapped to a 7.4-cM region of rice chromosome 11 containing the Xa21 gene, which confers resistance to the pathogen Xanthomonas oryzae pv oryzae. Three markers, RG103, 248 and 818, co-segregated with Xa21 in a population of 1141 plants. Multiple copies of all marker loci were present within the region that was introgressed from Oryza longistaminata into O. sativa. The marker loci were cloned and primers were designed that defined sequence-tagged sites. Physical mapping of the three tightly linked central markers revealed that RG103, the marker that hybridizes to the Xa21 gene, resides on a separate DNA fragment from the other two markers.Disclaimer: Names are necessary to report factually on available data; however, the USDA neither guarantees nor warrants the standard of the product, and the use of the name by USDA implies no approval of the product to the exclusion of others that may also be suitable. 相似文献
67.
Ana Cristina Gomes da Cunha Manuel Fernandes Ferreira 《Plant Cell, Tissue and Organ Culture》1996,47(1):1-8
The effects of plant growth regulators (PGR) on calli induction, morphogenesis and somatic embryogenesis of flax were studied. The organogenic and callus formation capacity were assessed for different types of source explants. Root and shoot explants were equally good material for calli production but the former produced calli without shoot regeneration capacity. Under the experimental conditions tested, 2,4-dichlorophenoxyacetic acid (2,4-D) + zeatin was the most efficient PGR combination on calli induction and biomass production. The calli were green but with no rhizogenic capacity. In contrast, and at similar concentrations, indole-3-butyric acid (IBA) + kinetin induced white or pale green friable calli with a good root regeneration capacity (60%). A factorial experiment with different combinations of 2,4-D + zeatin + gibberellic acid (GA3) levels revealed that the direction of explant differentiation was determined by specific PGR interactions and concentrations. The results from these experiments revealed that the morphogenetic pathway (shoot versus root differentiation) can be manipulated on flax explants by raising the 2,4-D level from 0.05 to 3.2 mg l?1 in the induction medium. The induction and development of somatic embryos from flax explants was possible in a range of 2,4-D + zeatin concentrations surrounding 0.4 mg l?1 2,4-D and 1.6 mg l?1 zeatin, the most efficient growth regulator combination. 相似文献
68.
Phylogeny and substitution rates of angiosperm actin genes 总被引:13,自引:1,他引:12
Forty-four actin genes from five angiosperm species were PCR-amplified,
cloned, and sequenced. Phylogenetic analysis of 34 of these actins, along
with those previously published, indicates that angiosperm actin genes are
monophyletic and underwent several duplications during evolution.
Orthologues have been identified between Solanaceae species, as well as
between Solanaceae species and soybean. These sequences were used to
calculate nucleotide substitution rates. The synonymous rate (6.96 x 10(-9)
substitutions/site/year) is similar to that of other nuclear protein-coding
genes, but the nonsynonymous rate (0.19 x 10(-9) substitutions/site/year)
is 6-19 times higher than that of mammalian actin genes. Relative rate
tests indicate that actin genes are evolving at similar rates in monocots
and in dicots. Evidence is also presented that some members of the maize
actin multigene family have been involved in gene conversion events, that
the potato genome contains 24 +/- 12 actin genes, and that potato and
tomato diverged 11.6 +/- 3.6 MYA.
相似文献
69.
The operational stability of the Candida rugosa lipase immobilized in a hydrophilic polyurethane foam was evaluated in consecutive batches for the glycerolysis of olive oil in n-hexane, aimed at the production of monoglycerides.Glycerol controlled the glycerolysis in the system under study, since it is both a substrate and a powerful water binder that reduces the water activity of the reaction medium and of the microenvironment. Two sets of experiments were carried out under different glycerol/triglyceride ratios. After 345 hours of consecutive 23 hours batches no lipase inactivation was observed.List of Symbols aw
thermodynamic activity of water
- DG
diglyceride (s)
- FAME
fatty acid methyl ester (s)
- FFA
free fatty acid (s)
- FID
flame ionization detector Gly glycerol
- MG
monoglyceride (s)
- TG
triglyceride (s)
- TLC
thin layer chromatography
The authors are grateful to Profs. P. Adlercreutz, Technical University of Lund, Sweden, and J.M.S. Cabral, Instituto Superior Técnico, Lisbon, Portugal, for inspiring discussions and advice, to Prof. Helena Pereira, Instituto Superior de Agronomia (ISA), Lisbon, Portugal, for the use of GC equipment and to Mrs. Marlene Dionísio, ISA, for invaluable help with some of the experimental work. 相似文献
70.
M. H. L. Ribeiro D. M. F. Prazeres J. M. S. Cabral M. M. R. da Fonseca 《Bioprocess and biosystems engineering》1995,12(1-2):95-102
l-tryptophan was produced froml-serine and indole by immobilized Escherichia coli cells in organic-aqueous systems. Selective adsorption was the method chosen to enable both product separation andl-serine reutilization. Amongst various adsorbents tested activated carbons and neutral polymeric resins (XAD-4 and XAD-7) showed good performance. The neutral resins could selectively concentrate thel-tryptophan from dilute aqueous solutions and adsorbed only 5% of the unconvertedl-serine. High separation factors (l-tryptophan/l-serine and indole/l-tryptophan) were obtained with these adsorbents. Despite a lower capacity, the XAD-7 resin had the advantage of desorbingl-tryptophan with basic or acidic solutions, while organic solvents were required to desorb, at the same concentration levels, this compound from XAD-4.In a packed bed column filled with XAD-4 resin or activated carbon, totall-tryptophan adsorption and recovery were achieved at linear velocities up to 5.0 cm/min and 3.2 cm/min respectively. Successive sorbent reutilization, following continuous sorption and elution steps, was carried out in packed bed columns with the neutral resins and activated carbon.Thel-form of tryptophan, after crystallization, was identified by HPTLC.List of Symbols HPLC
High Performance Liquid Chromatography
- HPTLC
High Performance Thin Layer Chromatography
- Trp
tryptophan
- Ser
Serine
-
A
amount of sorbent(g)
-
c
equilibrium solute concentration in the aqueous phase (g/dm3)
-
c
i
initial (before adding the sorbent) liquid phase concentration (g/dm3)
-
C
T
tryptophan concentration in the inlet solution (g/dm3)
-
C
To
tryptophan concentration in the outlet solution (g/dm3)
-
E
z
axial dispersion coefficient (m2/s)
-
k
experimental constant (Eq. 1, 2 and 3)
-
K
1
rate constant of adsorption (min–1)
-
L
column length(m)
-
n
experimental constant (eq. 1, 2 and 3)
-
q
equilibrium solid phase concentration (g solute/g sorbent)
-
q
max
maximum capacity of sorbent (g solute/g sorbent)
-
t
time(s)
-
v
liquid velocity (m/s)
-
V
volume of liquid phase(dm3)
-
V
e
eluted volume(dm3)
-
V
r
volume needed to saturate the column (dm3) 相似文献