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81.
82.
The complete gene coding for human neutrophilactivating protein-1/interleukin-8 was synthesized using a semi-chemical semi-enzymatic method. The synthetic gene was then overexpressed in Escherichia coli under the temperature-regulated control of the P_RP_L tandem promoters. As determined by SDS-PAGE and densitometry, the overexpressed protein comprised up to 18.5% and 10.9% of the total soluble protein in E. coli cells grown in shake flasks and in batch fermentation, respectively. The recombinant NAP-1/IL-8 was then purified to>95% homogeneity by gel filtration and cation exchange chromatography. The purified protein appeared as a single band on the SDS-PAGE gel and possessed potent chemotactic activity in the concentration of <10 ng/ml, as assayed by the agarose plate method. An early skin reactivity was also observed when the pure NAP-1/IL-8 was injected subcutaneously into the rabbits. The N-terminal 36 amino acid sequence of the recombinant NAP1/IL-8 was determined using the Edman method and was sho 相似文献
83.
The proline biosynthetic pathway and Pro genes in Saccharomyces cerevisiae have just begun to be studied recently. In our laboratory, Pro2 gene of S. cerevisiae had been cloned in yeast. As described in this paper, yeast Pro3 gene was also cloned, which can complement yeast Pro3 mutants, and be expressed efficiently in E. coli. The high activities of this gene product, L-pyrroline-5-carboxylate (P5C) reductase, can be detected in both organisms. The activity of the Pro3 gene product in multiple copy plasmids is not higher than that of single copy genes in chromosomes in both yeast and E. coll. The preliminary characterization of the gene is also reported. 相似文献
84.
Measurement of the degree of crystallinity of the polymer matrix in a composite is complicated by the presence of the reinforcing additive. This is particularly the case in APC-2 in which as much as 70% can be carbon fibre. A First Law procedure, developed for determining the degree of crystallinity of PEEK, which involves direct measurement of the enthalpy changes associated with melting, crystallization and heat capacity changes, has found to be an effective method for the determination of the crystallinity of the PEEK matrix. The procedure has been applied to carbon fibre and glass fibre PEEK composites. 相似文献
85.
The molar enthalpies of the solid–solid and solid–liquid phase transitions were determined by differential scanning calorimetry
for pure TbCl3 and KTb2Cl7, RbTb2Cl7, CsTb2Cl7, K3TbCl6, Rb3TbCl6 and Cs3TbCl6 compounds. Both types of compounds, i.e. M3TbCl6 and MTb2Cl7 (M=K, Rb, Cs) melt congruently and show additionally a solid–solid phase transition with a corresponding enthalpy Δtrs
H
0 of 6.1, 7.6 and 7.0 kJ mol–1 for potassium, rubidium and caesium M3TbCl6 compounds andΔtrs
H
0 of 17.1 (rubidium) and of 12.1 and 10.9 kJ mol–1 (caesium) for MTb2Cl7 compounds, respectively. The enthalpies of fusion were measured for all the above compounds with the exception of Rb3TbCl6 and Cs3TbCl6. The heat capacities of the solid and liquid compounds have been determined by differential scanning calorimetry (DSC) in
the temperature range 300–1100 K. The experimental heat capacity strongly increases in the vicinity of a phase transition,
but varies smoothly in the temperature ranges excluding these transformations. C
p data were fitted by an equation, which provided a satisfactory representation up to the temperatures of C
p discontinuity. The measured heat capacities were checked for consistency by calculating the enthalpy of formation of the
liquid phase, which had been previously measured. The results obtained agreed satisfactorily with these experimental data.
This revised version was published online in August 2006 with corrections to the Cover Date. 相似文献
86.
Functional expression of a β-d-1,4 glucanase-encoding gene (egl1) from a filamentous fungus was achieved in both Escherichia coli and Saccharomyces cerevisiae using a modified version of pRS413. Optimal activity of the E. coli-expressed enzyme was found at incubation temperatures of 60°C, whereas the enzyme activity was optimal at 40°C when expressed
by S. cerevisiae. Enzyme activity at different pH levels was similar for both bacteria and yeast, being highest at 5.0. Yeast expression resulted
in a highly glycosylated protein of approx 60 kDa, compared to bacterial expression, which resulted in a protein of 30 kDa.
The hyperglycosylated protein had reduced enzyme activity, indicating that E. coli is a preferred vehicle for production scale-up. 相似文献
87.
Yujie Wang Lifang Ruan Wai-Hung Lo Hong Chua Hoi-Fu Yu 《Applied biochemistry and biotechnology》2006,132(1-3):1015-1022
Polyhydroxyalkanoates (PHAs) are polyesters of hydroxyalkanoates synthesized by numerous bacteria as intracellular carbon
and energy storage compounds and accumulated as granules in the cytoplasm of cells. In this work, we constructed two recombinant
plasmids, pBE2C1, and pBE2C1AB, containing one or two PHA synthse, genes, respectively. The two plasmids were inserted into
Bacillus subtilis DB104 to generate modified strains, B. subtilis/pBE2C1 and B. subtilis/pBE2C1AB. The two recombinants strains were subjected to fermentation and showed PHA accumulation, the first reported example
of mcl-PHA production in B. subtilis. Gas Chromatography analysis identified the compound produced by B. subtilis/pBE2C1 to be a hydroxydecanoate-co-hydroxydodecanoate (HD-co-HDD) polymer whereas that produced by B. subtilis/pBE2C1AB was a hydroxybutyrate-co-hydroxyde-canoate-co-hydroxydodecanoate (HB-HD-HDD) polymer. 相似文献
88.
Bacillus cellulyticus K-12 Avicelase (Avicelase I; EC 3.2.1.4) gene (ace A) has been cloned in Escherichia coli by using the vector pT7T3U19 and HindIII-HindIII libraries of the chromosomal inserts. The libraries were screened for the expression of avicelase by monitoring the immunoreaction
of the antiavicelase (immunoscreening). Positive clones (Ac-3, Ac-5, and Ac-7) contained the identical 3.5-kb HindIII fragment as determined by restriction mapping and Southern hybridization, and expressed avicelase efficiently and constitutively
using its own promoter in the heterologous host. From the immunoblotting analysis, a polypeptide that showed a carboxymethylcellulase
(CMCase) activity with an M
r
, of 64,000 was detected. The recombinant endo 1,4-β-
d
-glucanase I was purified to homogeneity from an intracellular fraction of E. coli by DEAE-Toyopearl M650, Phenyl Toyoperal M650, and TSK gel HW50S chromatography. The enzyme had a monomeric structure, its
relative molecular mass being 65 kDa by gel filtration and 64 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
The pI was 5.3 and the optimal pH was 4.6, and the enzyme was stable at pH 4.0–10.5. The enzyme had a temperature optimum of 50°C
and was stable at 55°C for 48 h, and retained approx 20% of its activity after 30 min at 70°C. It showed high activity toward
carboxymethylcellulose (CMC) as well as p-nitrophenyl-β-d-cellobioside, 4-methylumbelliferyl cellobioside, Avicel, filter paper, and some cellooligosaccharides. K
m
values for CMC and Avicel were 7.6 and 85.2 mg/mL, respectively, whereas V
max values were 201 and 9.2 μmol · min−1 · mg−1, respectively. Cellotetraose (G4) was preferentially cleaved into cellobiose (G2) and cellopentaose (G5) was cleaved into
G2 + cellotriose (G3), whereas cellohexaose (G6) was cleaved into G4 + G2 and, to a lesser extent, into G3 + G3. G3 was not
cleaved at all. G2 was the main product of Avicel hydrolysis. G2 inhibited whereas Mg++ stimulated the activity of CMCase and Avicelase. Hydrolysis of CMC took place with a rapid decrease in viscosity but a slow
liberation of reducing sugars. Based on these results, it appeared that the cellulase should be regarded as endo type, although
it hydrolyzed Avicel. 相似文献
89.
The areas of the fusion and crystallization peaks of K3TaF8 and K3TaOF6 have been measured using the DSC mode of the high-temperature calorimeter (SETARAM 1800 K). On the basis of these quantities and the temperature dependence of the used calorimetric method sensitivity, the values of the enthalpy of fusion of K3TaF8 at temperature of fusion 1039 K: ΔfusHm(K3TaF8; 1039 K) = (52 ± 2) kJ mol−1 and of K3TaOF6 at temperature of fusion 1055 K: ΔfusHm(K3TaOF6; 1055 K) = (62 ± 3) kJ mol−1 have been determined. 相似文献
90.
Comment on papers by K. Shanahan that propose to explain anomalous heat generated by cold fusion 总被引:1,自引:0,他引:1
Edmund Storms 《Thermochimica Acta》2006,441(2):207-209
Dr. Shanahan has published two papers (Thermochim. Acta 428 (2005) 207, Thermochim. Acta 382 (2002) 95) in which he argues that excess heat claimed to be produced by cold fusion is actually caused by errors in heat measurement. In particular, he proposes that unrecognized changes in the calibration constant are produced by changes in the locations where heat is being generated within the electrolytic cell over the duration of the measurement. Because these papers may lend unwarranted support to rejection of cold fusion claims, these erroneous arguments used by Shanahan need to be answered. 相似文献