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91.
Stephan-Olav Wenk Jochen Kruip 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2000,737(1-2)
New porous materials have been tested for their potential to speed up purification of membrane proteins. As an example the purification of photosystem I, a light-driven electron pump from the cyanobacterium Synechocystis PCC6803, was optimized. The combination of two HPLC steps (an anion-exchange chromatography followed by a hydrophobic interaction chromatography) yields homogeneous monomeric or trimeric photosystem I as determined by gel filtration and gel electrophoresis. In comparison to traditional purification schemes our method is at least three-times faster and allows for easy scale-up. 相似文献
92.
A RelC deletion mutant, KO-100, of Streptomyces coelicolor A3(2) has been isolated from a collection of spontaneous thiostrepton-resistant mutants. KO-100 grows as vigorously as the
parent strain and possesses a 6-bp deletion within the rplK, previously termed relC. When the wild-type rplK gene was propagated on a low-copy-number vector in mutant KO-100, the ability to produce ppGpp, actinorhodin and undecylprodigiosin,
which had been lost in the RelC mutant, was completely restored. Allele replacement by gene homogenotization demonstrated
that the RelC mutation is responsible for the resistance to thiostrepton and the inactivation of ppGpp, actinorhodin and undecylprodigiosin
production. Western blotting showed that ribosomes from the RelC mutant KO-100 contain only one-eighth the amount of L11 protein
found in ribosomes of the parent strain. The impairment of antibiotic production in KO-100 could be rescued by the introduction
of mutations that confer resistance to streptomycin (str), which result in alteration of Lys-88 in ribosomal protein S12 to Glu or Arg. No accompanying restoration of ppGpp synthesis
was detected in these RelC str double mutants.
Received: 12 May 1997 / Accepted: 22 July 1997 相似文献
93.
94.
JOHN A. KLOETZEL 《The Journal of eukaryotic microbiology》1991,38(4):392-401
Morphogenesis of the ciliate cortex has been viewed as an attractive model system for studying the mechanisms behind the ordered assembly of subcellular structure. Based on the assumption that identifying protein components of the cortex would facilitate the study of cortical assembly, I have produced a number of monoclonal antibodies directed against components of the cortex of Euplotes aediculatus. Several of these antibodies react with the proteins comprising the alveolar plates. These thin polygonal scales, each enclosed within a flattened membranous sac (alveolus) just beneath the cell membrane, tightly abut in a confluent monolayer that appears to lend form and rigidity to the Euplotes cell cortex. Reactivity and specificity of these monoclonal antibodies for the alveolar plates was shown by immunofluorescence staining of whole-cell preparations and of cryosections, and by immuno-gold staining of thin sections by electron microscopy. On immunoblots of SDS-PAGE separated whole-cell extracts, the plate proteins are revealed as two to three closely spaced bands centered at an Mr of 97 kDa, and a larger relative at 125 kDa. Comparative peptide mapping reveals that the members of the 97-kDa protein cluster are closely related. However, the 125-kDa polypeptide varies significantly from the 97-kDa members, and hence is not likely a synthetic precursor. Because bands of these Mr values are prominent in Coomassie blue-stained gels of whole-cell extracts, and are greatly enriched in purified cortical preparations, they likely represent the major proteins comprising the alveolar plates of E. aediculatus. I have proposed the name platein for this family of proteins. 相似文献
95.
The helicase superfamily 2 (SF2) proteins are involved in essentially every step in DNA and RNA metabolism. The radD (yejH) gene, which belongs to SF2, plays an important role in DNA repair. The RadD protein includes all seven conserved SF2 motifs and has shown ATPase activity. Here, we first reported the structure of RadD from Escherichia coli containing two RecA-like domains, a zinc finger motif, and a C-terminal domain. Based on the structure of RadD and other SF2 proteins, we then built a model of the RedD-ATP complex. 相似文献
96.
Seed protein profiles of 24 wild and cultivated taxa of Pisum have been compared by sodium dodecyl sulphate polyacrylamide gel electrophoresis. No consistent differences were detected either among wild taxa or between wild and cultivated taxa. This shows that Pisum forms a single-species complex on the basis of seed protein profiles. 相似文献
97.
Summary It is widely accepted that seed storage proteins accumulate only in cells which have entered the cell expansion phase and do not continue to divide. Here we present data indicating that the accumulation of storage globulins in tobacco begins already during early embryogenesis in a period of sustained mitotic activity. Western blot analysis revealed that polypeptides of the legumin-like 12S globulins (Mr 60000, 40000, 20000) appear at mid/late globular stage, whereas the vicilin-like 7S globulin (Mr 50000) follows during the transition from heart to torpedo stage. The accumulation of legumin-like polypeptides begins first in the endosperm during the mid globular stage followed in the embryo-suspensor complex during the heart-shaped stage. The vicilin-related fraction appears first in the endosperm and three days later in the embryo. Examination of individual cells from squash preparations revealed that protein bodies are not confined to intermitotic cells, but are also present in cells undergoing mitosis. Protein bodies of dividing cells situated outside the mitotic apparatus are not metabolized during cytokinesis. The only cell type which loses its protein bodies completely prior to the first mitotic division is the primary hypophysis cell. Our finding that storage proteins can occur in dividing cells independent of their origin and developmental capacity indicates that the cell expansion hypothesis of storage protein accumulation has to be revised. 相似文献
98.
Using a histochemical technique, we found that in rat embryos heparin-binding sites are localized within ventricular regions of the neural tube. The highest intensity of the heparin-binding activity was observed in the membranes of migrating nerve cells. Heparin-binding membrane-associated proteins were isolated and purified from the brains of newborn rats; molecular masses of two such proteins were measured (19 and 28 kdalton). The level of affinity for binding of heparan sulfate to the purified proteins was characterized by equilibrium constants of 1.7 · 10-3 and 6.7 · 10-3. Binding of heparan sulfate to the above proteins was more intensive at low ion force and pH values within the 3.0 to 4.0 range and about 6.0. 相似文献
99.
H. Vis C. M. Dobson C. V. Robinson 《Protein science : a publication of the Protein Society》1999,8(6):1368-1370
Nanoflow electrospray mass spectrometry was used to monitor the formation of protein heterodimers of HU proteins from Bacillus stearothermophilus and Bacillus subtilis. This has enabled us to analyze both thermodynamic and kinetic features associated with the dissociation of homodimeric HU proteins. The results obtained correlate well with the kinetics of the protein dissociation process and the free energy difference between homo- and heterodimeric species anticipated from other studies. We suggest that this approach will have general applicability in studying protein association and dissociation under near-equilibrium conditions and will be relevant to a wide range of biological systems. 相似文献
100.