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1.
The distribution of Escherichia coli recA protein bound to duplex DNA with single-stranded ends 总被引:2,自引:0,他引:2
A short single-stranded tail on one end of an otherwise duplex DNA molecule enables recA protein, in the presence of ATP and MgCl2, to form a complex with the DNA which extends into the duplex portion of the molecule. Nuclease protection studies at a concentration of MgCl2 which permits homologous pairing showed that cleavage by restriction endonucleases at sites throughout the duplex region was inhibited, whereas digestion by DNase I was not affected. These results indicate that recA protein binds to the duplex portion of tailed DNA allowing access by DNase I to a random sample of the many sites at which it cleaves, but providing limited protection of the relatively rare restriction sites. Electron microscopy revealed that the recA nucleoprotein complex with duplex DNA is indeed a segmented or interrupted filament that, with time, extends further from the single-stranded tail into the duplex region. recA protein binding extended into the duplex region more rapidly for duplexes with 5' tails than for those with 3' tails. These observations show that recA protein translocates from a single-stranded region into duplex DNA in the form of a segmented filament by a mechanism that is not strongly polarized. 相似文献
2.
T Yoshikawa W Flory D H Giamalva L P Ruhr D F Church W A Pryor 《Biochemistry international》1987,15(1):147-151
Male Sprague-Dawley rats were treated ip with beta-naphthoflavone (40 mg/kg/day) in corn oil or in DMSO for three days. Diphenaldehyde (90 mg/kg in DMSO) was injected ip 24 hr after pretreatment. The increase in the levels of aspartate aminotransferase, alanine aminotransferase, sorbitol dehydrogenase, gamma glutamyl transpeptidase, and lactate dehydrogenase was significantly lower in rats pretreated with BNF. This suggests that the BNF-induced P-450 isozyme systems have a protective effect against the acute hepatotoxicity of diphenaldehyde. 相似文献
3.
Peter Kaastrup René J. M. Stet Albert J. Tigchelaar Egbert Egberts Willem B. van Muiswinkel 《Immunogenetics》1989,30(4):284-290
In this study, experiments are described that were designed to investigate whether fish have an immune regulatory systems
similar to the major histocompatibility complex (MHC) in higher vertebrate species. From combinations of gynogenetic carp
showing either slow of fast rejection of skin transplants, the latter were chosen for alloantiserum production by hyperimmunization
with peripheral blood leucocytes. The resulting alloantisera were analyzed for hemagglutinating reactivity with gynogenetic
siblings and proved to be operationally monoscpecific in absorption experiments. The serologically determined carp erythrocyte
specificites were shown to correspond to two codominantly expressed allelic products of a single locus and were designated
K1 and K2, respectively. Flow cytometer analysis revealed that the same products are also present on leucocytes from peripheral
blood, thymus, spleen, and pronephros.K1-andK2-homozygous second-generation gynogenetic siblings were used to study the histocompatibiligy nature of the K locus products.
Skin transplants between K-allogeneic gynogenetic siblings were rejected significantly faster than within K-syngeneic combinations.
Taken together, these data suggest that theK locus incorporates MHC class I-like characteristics. 相似文献
4.
T Yoshikawa W Flory D H Giamalva L P Ruhr D F Church W A Pryor 《Biochemistry international》1988,16(5):929-934
Male Sprague-Dawley rats were pretreated i.p. with corn oil or DMSO (1.5 ml/kg/day), or with beta-naphthoflavone (BNF, 40 mg/kg/day) in corn oil or DMSO for 3 days. 1-Nitropyrene (1-NP, 150 mg/kg) in DMSO was injected i.p. 24 hr after pretreatment. A significant increase in the levels of several serum enzymes was seen in rats pretreated with corn oil alone 24 hr after 1-NP injection. The increase in enzyme activities was significantly reduced by a 3-day pretreatment with DMSO or BNF. 相似文献
5.
A series of ring-substituted 3-phenylpropenes has been examined as mechanism-based inhibitors for the copper protein dopamine beta-hydroxylase. p-HO-, p-CH3O-, m-HO-, m-CH3O-, p-Br-, and p-CN-substituted phenylpropenes all inactivate the enzyme under turnover conditions, requiring ascorbate and oxygen. Replacement of the benzylic hydrogens in 3-(p-hydroxyphenyl)propene with deuterium results in a kinetic isotope effect of 2.0 on kinact/KO2 but in no effect on the partition ratio, Vmax/kinact, consistent with a stepwise mechanism for hydrogen abstraction and oxygen insertion. The partition ratio is unchanged in the pH range from 4.5 to 7.1. Determination of the kinetics of inactivation and the partition ratios for each of these ring-substituted phenylpropenes has allowed determination of the respective V/KO2 values. A linear free energy plot of these values as a function of sigma+ gives a rho value of -1.2, while the partition ratios show only a slight decrease upon going electron-withdrawing groups. The results are consistent with a mechanism for dopamine beta-hydroxylase in which a hydrogen atom is abstracted to form a benzylic radical, which then partitions between hydroxylation and enzyme inactivation. 相似文献
6.
Kinetics of the activation of rat liver pyruvate kinase by fructose 1,6-disphosphate and methods for characterizing hysteretic transitions
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H. Olin Spivey Wayne Flory Benigno D. Peczon John P. Chandler Roger E. Koeppe 《The Biochemical journal》1974,141(1):119-125
1. Kinetics of fructose 1,6-diphosphate activation of liver pyruvate kinase type I inhibited with MgATP and l-alanine are described as a function of enzyme and fructose 1,6-diphosphate concentrations. These results can be explained by a single pseudo-first-order transition of the enzyme into an active form, independent of the enzyme concentration. This rate constant, k(app.)=0.24s(-1) with 0.02mm-fructose 1,6-diphosphate (t(0.9) approximately 10s where t(0.9) is the time for 90% conversion), is an increasing function of fructose 1,6-diphosphate concentration far beyond that needed to maximally activate enzyme equilibrated with fructose 1,6-diphosphate (about 20mum). 2. The model equations are best analysed with numerical techniques which are described. These techniques are useful in studying similar slow transients frequently observed in stopped-flow studies of enzymes. 3. Shorter transients (t(0.9)=0.5-1.5s) were observed in the kinetic response of the enzyme to the addition of MgATP or phosphoenolpyruvate, but were not further characterized. 相似文献
7.
Molar Kerr contants mK have been determined for N-methylacetamide, N-ethyl-acatamide, N-methylpropionamide, N-acetylglycine-N′-methylamide, N-acetyl-L -ala-nine-N′-methylamide, and N-acetyl-L -leucine-N′-methylamide from electric birefringence measurements on dilute solutions in dioxane. The resultes are compared with configurational averatges 〈mK〉 calculated on the basis of conformational energies. Values ofmK calculated for various fixed confromations on the peptides vary over a wide range depending on the choice of conformation; hece, the configurational averages 〈mK〉 are sensitive to smalll changes in the conformational enerey. Kerr constants are calculated for random coiled oligoglycine and oligoalanine peptides as functions of chain length. The Kerr constant is an especially acute index of the average conformation. 相似文献
8.
9.
Michael Steinert Manfred Ott P. Christian Lück Egbert Tannich Jörg Hacker 《FEMS microbiology ecology》1994,15(3-4):299-308
Abstract Legionella pneumophila strains isolated from different sources were tested for their host range in the protists Acanthamoeba castellanii, Hartmannella vermiformis and Entamoeba histolytica . It has been shown that A. castellanii and H. vermiformis but not E. histolytica support the intracellular replication of L. pneumophila . Furthermore it could be demonstrated that in vivo virulence in the guinea pig and the intracellular growth in U937 cells coincides with the capability to replicate intracellularly in A. castellanii at 37°C. The infectivity of L. pneumophila that had sustained a 48 hours nutrient deprivation was not significantly different from that of legionellae grown to log-phase on BCYE plates. In contrast the nutrient limitation on A. castellanii increased the amount of intracellular legionellae at the beginning of infection. An initial opsonin independent attachement stage of legionellae to U937 cells was demonstrated by scanning electron microscopy. In contrast, L. pneumophila's capability of stable or long term attachmennt to A. castellanii was shown to be inefficient. 相似文献
10.
Egbert J. Boekema Arjen F. Boonstra Jan P. Dekker Matthias Rögner 《Journal of bioenergetics and biomembranes》1994,26(1):17-29
Electron microscopy (EM) in combination with image analysis is a powerful technique to study protein structure at low- and high resolution. Since electron micrographs of biological objects are very noisy, substantial improvement of image quality can be obtained by averaging individual projections. Crystallographic and noncrystallographic averaging methods are available and have been applied to study projections of the large protein complexes embedded in photosynthetic membranes from cyanobacteria and higher plants. Results of EM on monomeric and trimeric Photosystem I complexes, on monomeric and dimeric Photosystem II complexes, and on the monomeric cytochromeb6/f complex are discussed. 相似文献