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Ty1 is a retrotransposon of the yeast Saccharomyces cerevisiae whose transposition at new locations in the host genome is activated by stress conditions, such as exposure to UV light, X-rays, nitrogen starvation. In this communication, we supply evidence that cooling for 2 h at +4 °C followed by freezing for 1 h at −10 °C and 16 h at −20 °C also increased Ty1 transposition. The mobility of Ty1 was induced by cooling at slow rates (3 °C/min) and the accumulation of trehalose inside cells or the cooling at high rates (100 °C/min) inhibited significantly the induction of the transposition. The freeze-induced Ty1 transposition did not occur in mitochondrial mutants (rho−) and in cells with disrupted SCO1 gene (Δsco1 cells) evidencing that the Ty1 transposition induced by cooling depends on the mitochondrial oxidative phosphorylation. We also found that the freeze induced Ty1 transposition is associated with increased synthesis and accumulation of superoxide anions (O−2) into the cells. Accumulation of O−2 and activation of Ty1 transposition were not observed after cooling of cells with compromised mitochondrial functions (rho−, Δsco1), or in cells pretreated with O−2 scavengers. It is concluded that (i) elevated levels of reactive oxygen species (ROS) have a key role in activation the transposition of Ty1 retrotransposon in yeast cells undergoing freezing and (ii) given the deleterious effect of increased ROS levels on cells, special precautions should be taken to avoid ROS production and accumulation during cryopreservation procedures. 相似文献
105.
Devred F Tsvetkov PO Barbier P Allegro D Horwitz SB Makarov AA Peyrot V 《FEBS letters》2008,582(17):2484-2488
Microtubule (MT) dynamic instability is tightly regulated by stabilizing and destabilizing proteins, the latter being exemplified by stathmin/Op18, a protein known to destabilize MTs. Studies in cells have indicated that the level of stathmin expression modifies the cytotoxicity of antimicrotubule drugs, such as vinblastine (VLB). Using isothermal titration calorimetry and analytical ultracentrifugation, we show that VLB increases the affinity of stathmin for tubulin 50-fold (and vice versa). These results are the first biochemical evidence of the direct relationship between stathmin and an antimitotic drug, and reveal a new mechanism of action for VLB. 相似文献
106.
The effect of various toxic compounds (phenol, gasoline, detergents, halogenated benzenes, and copper salts) on the activity and multiple forms of acid DNase was studied in the liver of the widely occurring freshwater snail species Viviparus viviparus L. Characteristic variations in the specific activity and isoform pattern of the enzyme were revealed depending on pollutant concentration and exposure time. It was shown that the pattern of DNase isoforms in V. viviparus could be an index of water pollution. 相似文献
107.
Binding of ATP to heat shock protein 90: evidence for an ATP-binding site in the C-terminal domain. 总被引:4,自引:0,他引:4
Cyrille Garnier Daniel Lafitte Philipp O Tsvetkov Pascale Barbier Jocelyne Leclerc-Devin Jean-Marc Millot Claudette Briand Alexander A Makarov Maria G Catelli Vincent Peyrot 《The Journal of biological chemistry》2002,277(14):12208-12214
The presence of a nucleotide binding site on hsp90 was very controversial until x-ray structure of the hsp90 N-terminal domain, showing a nonconventional nucleotide binding site, appeared. A recent study suggested that the hsp90 C-terminal domain also binds ATP (Marcu, M. G., Chadli, A., Bouhouche, I., Catelli, M. G., and Neckers, L. M. (2000) J. Biol. Chem. 275, 37181-37186). In this paper, the interactions of ATP with native hsp90 and its recombinant N-terminal (positions 1-221) and C-terminal (positions 446-728) domains were studied by isothermal titration calorimetry, scanning differential calorimetry, and fluorescence spectroscopy. Results clearly demonstrate that hsp90 possesses a second ATP-binding site located on the C-terminal part of the protein. The association constant between this domain of hsp90 and ATP-Mg and a comparison with the binding constant on the full-length protein are reported for the first time. Secondary structure prediction revealed motifs compatible with a Rossmann fold in the C-terminal part of hsp90. It is proposed that this potential Rossmann fold may constitute the C-terminal ATP-binding site. This work also suggests allosteric interaction between N- and C-terminal domains of hsp90. 相似文献
108.
A. V. Deryugina L. V. Oshevenskiy M. N. Talamanova A. I. Tsvetkov M. A. Shabalin M. Yu. Glyavin V. N. Krylov 《Biophysics》2017,62(6):914-918
The effects of electromagnetic radiation of a gyrotron with an operating frequency of 263 GHz on rat erythrocyte electrophoretic mobility, malonic dialdehyde, ATP, and 2,3-diphosphoglycerate concentration have been studied in vitro. The blood was irradiated under continuous and pulsed gyrotron emission modes with the estimated incident energy flux on an object from 0.1 to 20 mW/cm2. It has been established that the studied erythrocyte characteristics changed in different directions depending on the mode and the incident energy flux: the erythrocyte electrophoretic mobility rate and ATP concentration decreased, while the malonic dialdehyde and 2,3-diphosphoglycerate concentration increased at a low incident energy flux and continuous exposure mode; all studied parameters increased under the pulsed exposure mode. An incident energy flux increase caused a reduction of these effects. 相似文献
109.
Intrinsically unstructured proteins (IUPs), also known as natively unfolded proteins, lack well-defined secondary and tertiary structure under physiological conditions. In recent years, growing experimental and theoretical evidence has accumulated, indicating that many entire proteins and protein sequences are unstructured under physiological conditions, and that they play significant roles in diverse cellular processes. Bioinformatic algorithms have been developed to identify such sequences in proteins for which structural data are lacking, but still generate substantial numbers of false positives and negatives. We describe here a simple and reliable in vitro assay for identifying IUP sequences based on their susceptibility to 20S proteasomal degradation. We show that 20S proteasomes digest IUP sequences, under conditions in which native, and even molten globule states, are resistant. Furthermore, we show that protein-protein interactions can protect IUPs against 20S proteasomal action. Taken together, our results thus suggest that the 20S proteasome degradation assay provides a powerful system for operational definition of IUPs. 相似文献
110.