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81.
Protein synthesis by synaptosomes from rat brain. Contribution by the intraterminal mitochondria 总被引:1,自引:0,他引:1
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A. G. Hernández 《The Biochemical journal》1974,142(1):7-17
(1) The characteristics of protein synthesis in microsomal and synaptosomal fractions from rat brain were examined. A high sensitivity to ribonuclease and to cycloheximide, and the need for the presence of pH5 enzymes distinguished protein synthesis in microsomal fractions from protein synthesis in synaptosomes. (2) Under various conditions of incubation synaptosomal fractions prepared in sucrose showed limited protein synthesis compared with synaptosomal fractions prepared by using Ficoll. Such discrepancies could not be attributed to: (i) animal age, (ii) the metabolic state of the synaptosomal fraction, (iii) the absence of bivalent cations in the incubation medium or (iv) the temperature. (3) Protein synthesis in synaptosomal fractions was inhibited 50-65% by cycloheximide, 38-50% by chloramphenicol, 95% by puromycin, 70% by azide and 40% by deoxyglucose; ribonuclease had only a negligible inhibitory effect. (4) As a first approximation to the localization of the protein-synthetic machinery present in the synaptosomal fraction, the distribution of enzymes and radioactivity in subfractions of prelabelled synaptosomes was determined after osmotic shock with water. Approximately 60% of the total protein synthesis in the synaptosomal fraction occurred in the intraterminal mitochondria. (5) Protein synthesis in the intraterminal mitochondria did not show any fundamental difference from synthesis in somatic mitochondria, with respect to inhibition by cycloheximide and chloramphenicol. (6) It was concluded that if extramitochondrial protein synthesis occurs in synaptosomes, it must be very low. 相似文献
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Summary The study of the cell division cycle by means of caffeine labelling inAllium roots, at 15° C, employing intact root and decapitated roots at several levels (0.5, 1.0, 1.5, 2.0, and 2.5 mm) has shown that the number of cycles developed by the cells is constant at each meristem level. This number and the durations of the cycles are not affected by the decapitation. It is suggested that the cell cycle is controlled in the meristematic cells by an intracellular programme which would be developed throughout the meristem.However, the larger the region decapitated is, the more decreases the growth rate of the roots. The removal of the root cap (about 0.5 mm) did not modify the rate of root growth, although it blocked the geotropic response. The quiescent center is proposed as a source of auxin controlling cell elongation. 相似文献
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86.
Characterization of the phage phi 29 protein p5 as a single-stranded DNA binding protein. Function in phi 29 DNA-protein p3 replication. 总被引:4,自引:2,他引:2
The phage phi 29 protein p5, required in vivo in the elongation step of phi 29 DNA replication, was highly purified from Escherichia coli cells harbouring a gene 5-containing plasmid and from phi 29-infected Bacillus subtilis. The protein was characterized as the gene 5 product by amino acid analysis and NH2-terminal sequence determination. The purified protein p5 was shown to bind to single-stranded DNA and to protect it against nuclease degradation. No effect of protein p5 was observed either on the formation of the p3-dAMP initiation complex or on the rate of elongation. However, protein p5 greatly stimulated phi 29 DNA-protein p3 replication at incubation times where the replication in the absence of p5 leveled off. 相似文献
87.
Maria F. Fernández Antonio Cobos Lorenzo Hoz Dr. Pablo E. Hernández Bernabé Sanz 《Current microbiology》1989,18(2):113-117
Operon fusions to the promoter of either theproA,proB, orproC genes of the proline biosynthetic pathway were obtained by the use of the Mu d1(Ap,lac) bacteriophage. These fusions were further stabilized by transformation with plasmid pGW600 containing the wildtype Mu repressor gene or by transduction with phage pSG1. The level of -galactosidase in the fusion strains was not affected by the presence of exogenously addedl-proline or high concentrations of NaCl in the growth medium. A Tn5 insertion nearproBA increased -galactosidase expression 140- to 200-fold in strains carrying theproA-lac andproB-lac fusions, but the level of this enzyme was unaltered in strains carrying theproC-lac fusion. The Tn5 insertion increased intracellular proline concentrations 8- to 10-fold, suggesting that mechanisms other than allosteric inhibition may regulate proline biosynthesis, but did not confer osmotolerance to cells growing in a medium with a high concentration of salt. 相似文献
88.
Subcellular localization of the major pneumococcal autolysin: a peculiar mechanism of secretion in Escherichia coli 总被引:9,自引:0,他引:9
E Díaz E García C Ascaso E Méndez R López J L García 《The Journal of biological chemistry》1989,264(2):1238-1244
The major pneumococcal autolysin (N-acetylmuramoyl-L-alanine amidase) has been localized in the cellular envelope of Streptococcus pneumoniae and Escherichia coli by using immunocytochemical labeling on ultrathin sections and whole-mounted cells. Cell fractionation experiments in E. coli confirmed the peripheral localization of the pneumococcal amidase and suggested that this enzyme is weakly bound to the outer face of the cytoplasmic membrane. This interaction does not depend on the presence of choline but represents an intrinsic property of the amidase. The autolysin, that is synthesized without any N-terminal signal sequence (García, P., García, J. L., García, E., and López, R. (1986) Gene (Amst.) 43, 265-272) was not processed during translocation. A new regulatory mechanism that might be specific for bacterial autolysins is discussed. 相似文献
89.
Activity of delta 5-3 beta-hydroxysteroid dehydrogenase coupled with steroid-delta 5-4-isomerase was demonstrated for the first time in the pancreas. The enzyme complex was assayed by measuring the conversion of pregnenolone to progesterone as well as of dehydroepiandrosterone to androstenedione and found to be localized primarily in the mitochondrial fraction of dog pancreas homogenates. The delta 5-3 beta-hydroxysteroid dehydrogenase used either NAD+ or NADP+ as co-substrates, although maximal activity was observed with NAD+. In phosphate buffer, pH 7.0 and 37 degrees C, the apparent Km values of the dehydrogenase were 6.54 +/- 0.7 microM for pregnenolone and 9.61 +/- 0.8 microM for NAD+. The apparent Vmax was determined as 0.82 +/- 0.02 nmol min-1 mg-1. Under the same conditions the Km values for dehydroepiandrosterone and NAD+ were 3.3 +/- 0.2 microM and 9.63 +/- 1.6 microM, respectively, and the apparent Vmax was 0.62 +/- 0.01 nmol min-1 mg-1. 相似文献
90.
C A Libreros-Minotta J P Pardo G Mendoza-Hernández J L Rendón 《Archives of biochemistry and biophysics》1992,298(1):247-253
Glutathione reductase (NAD(P)H:GSSG oxidoreductase EC 1.6.4.2.) was purified 1160-fold to homogeneity from the nonsulfurous purple bacteria Rhodospirillum rubrum (wild type). Specific activity of the pure preparation was 102 U/mg. The enzyme displayed a typical flavoprotein absorption spectrum with maxima at 274,365, and 459 nm and an absorbance ratio A280/A459 of 7.6. The amino acid analysis revealed an unusually high content of glycine and arginine residues. Titration of the enzyme with 5,5'-dithiobis(2-nitrobenzoic acid) showed a total of two free thiol groups per subunit, one of which is made accessible only under denaturing conditions. An isoelectric point of 5.2 was found for the native enzyme. Km values, determined at pH 7.5, were 6.1 and 90 microM for NADPH and GSSG, respectively. NADH was about 2% as active as NADPH as an electron donor. The enzyme's second choice in disulfide substrate was the mixed disulfide of coenzyme A and glutathione, for which the specific activity and Km values were 5.1 U/mg and 3.4 mM, respectively. A native molecular weight of 118,000 was found, while denaturing electrophoresis gave a value of 54,400 per subunit, thus suggesting that R. rubrum glutathione reductase exists as a dimeric protein. Other physicochemical constants of the enzyme, such as Stokes radius (4.2 nm) and sedimentation coefficient (5.71 S), were also consistent with a particle of 110,000. 相似文献