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21.
Ulrich Nowitzki Ralf Wyrich Peter Westhoff Katrin Henze Claus Schnarrenberger William Martin 《Plant molecular biology》1995,29(6):1279-1291
Exploiting the differential expression of genes for Calvin cycle enzymes in bundle-sheath and mesophyll cells of the C4 plant Sorghum bicolor L., we isolated via subtractive hybridization a molecular probe for the Calvin cycle enzyme d-ribulose-5-phosphate 3-epimerase (R5P3E) (EC 5.1.3.1), with the help of which several full-size cDNAs were isolated from spinach. Functional identity of the encoded mature subunit was shown by R5P3E activity found in affinity-purified glutatione S-transferase fusions expressed in Escherichia coli and by three-fold increase of R5P3E activity upon induction of E. coli overexpressing the spinach subunit under the control of the bacteriophage T7 promoter, demonstrating that we have cloned the first functional ribulose-5-phosphate 3-epimerase from any eukaryotic source. The chloroplast enzyme from spinach shares about 50% amino acid identity with its homologues from the Calvin cycle operons of the autotrophic purple bacteria Alcaligenes eutrophus and Rhodospirillum rubrum. A R5P3E-related eubacterial gene family was identified which arose through ancient duplications in prokaryotic chromosomes, three R5P3E-related genes of yet unknown function have persisted to the present within the E. coli genome. A gene phylogeny reveals that spinach R5P3E is more similar to eubacterial homologues than to the yeast sequence, suggesting a eubacterial origin for this plant nuclear gene.Abbreviations R5P3E
d-ribulose-5-phosphate 3-epimerase
- RPI
ribose-5-phosphate isomerase
- TKL
transketolase
- PRK
phosphoribulokinase
- GAPDH
glyceraldehyde-3-phosphate dehydrogenase
- FBP
fructose-1,6-bisphophatase
- FBP
fructose 1,6-bisphosphate
- G6PDH
glucose-6-phosphate dehydrogenase
- 6PGDH
6-phosphogluconate dehydrogenase
- OPPP
oxidative pentose phosphate pathway
- Rubisco
ribulose-1,5-bisphosphate carboxylase/oxygenase
- FBA
fructose-1,6-bisphophate aldolase
- IPTG
isopropyl -d-thiogalactoside
- GST
glutathione S-tranferase
- PBS
phosphate-buffered saline
- TPI
triosephosphate isomerase 相似文献
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24.
Beat W. Schfer Roland Wicki Dieter Engelkamp Marie-genevive Mattei Claus W. Heizmann 《Genomics》1995,25(3)
S100 proteins are low-molecular-weight calcium-binding proteins of the EF-hand superfamily and appear to be involved in the regulation of a number of cellular processes such as cell cycle progression and differentiation. More than 10 members of the S100 protein family have been described from human sources so far. We have now isolated a YAC clone from human chromosome 1q21, on which 9 different genes coding for S100 calcium-binding proteins could be localized. Moreover, we have mapped the gene coding for S100P to human chromosome 4p16 and thereby completed the chromosomal assignments of all known human S100 genes. The clustered organization of S100 genes in the 1q21 region allows us to introduce a new logical nomenclature for these genes, which is based on the physical arrangement on the chromosome. The new nomenclature should facilitate and further the understanding of this protein family and be easily expandable to other species. 相似文献
25.
Summary We have tested the acyl-transferase of penicillin producing strains and penicillin-less mutants of Penicillium chrysogenum for their ability to exchange the d--aminoadipic acid side chain of cephalosporin-C with phenoxyacetic acid. We found the reaction in both types of strains. 相似文献
26.
A method for the separation of the outer membrane (OM) from the cytoplasmic membrane (CM) of Acinetobacter calcoaceticus 69/V grown on different carbon sources is described. The contamination of the OM with CM was less than 10%. Independent of the carbon source, five protein bands with apparent molecular weights of 47 000, 33000, 21 000, 19 000 and 12 000 were found by solubilization at 37°C and six bands at 100°C (apparent Mr 53 000, 47 000, 38 000, 26 000, 21000, 12000). Three proteins were modifiable by heat. With the periodic acid-Schiff procedure the bands with apparent Mr of 33 000 and 12 000 were made visible. After growth on d,l-carnitine an additional two non-heat-modifiable protein bands with apparent Mr between 40 000 and 45 000 were detected. By cultivation on acetate and peptone as carbon source one additional band (Mr 15 000) from OM of cells could be found. 相似文献
27.
Changes in key regulatory enzymes of hepatic carbohydrate metabolism after glucose loading of starved rats 总被引:5,自引:0,他引:5
T H Claus F Nyfeler H A Muenkel M G Burns T Pate S J Pilkis 《Biochemical and biophysical research communications》1984,125(2):655-661
When glucose was given to starved rats there was an increase in both 6-phosphofructo 2-kinase and pyruvate kinase activity and a decrease in fructose 2,6-bisphosphatase activity 30 min and 60 min later. These changes were accompanied by an increase in glycogen deposition and by modest, but significant increases in fructose 2,6-bisphosphate levels at the same time. Metabolite measurements indicated that flux through 6-phosphofructo 1-kinase and pyruvate kinase were increased. These results suggest that although glycogen deposition may occur via the gluconeogenic pathway, glycolysis is activated at the same time by changes in the phosphorylation state of key regulatory enzymes as well as by the small rise in fructose 2,6-bisphosphate. 相似文献
28.
ACTIVITY AND ISOENZYME PATTERN OF LACTATE DEHYDROGENASE IN ASTROBLASTS CULTURED FROM BRAINS OF NEWBORN MICE 总被引:3,自引:0,他引:3
Lactate dehydrogenase activity and isoenzyme distribution was determined in primary cultures of astroblasts as a function of the culture period. The specific activity increased during this period with a peak value (1.91 ± 0.18μmol x min-1 x mg-1 cell protein) after 2 weeks in culture. The isoenzyme pattern changed during 3 weeks in culture towards a higher proportion of the H4 (LDH-1) isoenzyme which is analogous to the in vivo pattern. Omission of serum with or without dBcAMP (0.5 mM) in the culture medium during the third week of culture further enhanced this prominence of the H4 isoenzyme. The specific activity (1.58 × 0.06 μmol x min-1 x mg-1 cell protein) of cultures grown in the presence of 0.5 mM-dBcAMP and absence of serum was close to the activity in the adult brain. 相似文献
29.
Summary Fibroblasts of a patient with Bloom syndrome (GM-1492) were cultured in the presence of either mitomycin C, ethylmethanesulfonate, or 4-nitroquinoline-1-oxide, (4-NQ1-O) and sister chromatid exchange was determined. The mutagens enhanced the sister chromatid exchange rate to different degrees, 4-NQ1-O being the most potent substance. Bloom corrective factor, which is present in normal cell-conditioned culture medium, reduced the spontaneously increased SCE in Bloom syndrome cells by about 20 SCE per metaphase but failed to reduce the additional mutagen-induced SCE increase. These findings indicate that only spontaneously, but not mutagen-indeuced, SCE in Bloom syndrome fibroblasts can be decreased by the Bloom corrective factor. 相似文献
30.
A sensitive agarose diffusion method for the determination of α-amylase has been developed, using Reactone Red 2 B-amylopectin as the substrate. The logarithm of enzyme activity is linearly correlated with the diameters of the diffusion zones over a very extended range from 1 mU/ml to at least 1100 U/ml. The α-amylase activity in biological samples may be determined without dilution or pretreatment, and the test can be performed at any desired temperature between 4 and 45°C. The clear radial diffusion zones may be fixed, further enhancing the contrast to the bright red surrounding. 相似文献