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1.
Abstract A mutant strain of Schizosaccharomyces pombe lacking dipeptidyl aminopeptidase yspI was isolated from a strain already defective in aminopeptidase activity by means of a staining technique with the chromogenic substrate ala-pro-4-methoxy-β-naphthylamide to screen colonies for the absence of the enzyme. The defect segregated 2+ :2− in meiotic tetrads, indicating a single chromosomal gene mutation, which was shown to be recessive. Gene dosage experiments indicated that the mutation resides in the structural gene of dipeptidyl aminopeptidase yspI, dpa 1+ . The dpa 1+ gene was located on chromosome III by using l m- fluorophen-ylalanine-induced haploidization and mitotic analysis. dpa1 mutants did not show any obvious phenotype under a variety of conditions tested. 相似文献
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Sabrina Piombo Gode B. Calleja Bong Yul Yoo Byron F. Johnson 《Cell biochemistry and biophysics》1998,29(3):263-279
Distributions of rupture sites of fission yeast cells ruptured by glass beads have been related to a new morphometric analysis.
As shown previously (Johnson et al.,Cell Biophysics, 1995), ruptures were not randomly distributed nor was their distribution dictated by geometry, rather, ruptures at the extensile
end were related to cell length just as the rate of extension is related to cell length. The extension patterns of early log,
mid-log, late log, and stationary phase cells from suspension cultures were found to approximate the linear growth patterns
of Kubitschek and Clay (1986). The median length of cells was found to decline through the log phase in an unbalanced manner. 相似文献
5.
N. Arfman E. M. Watling W. Clement R. J. van Oosterwijk G. E. de Vries W. Harder M. M. Attwood L. Dijkhuizen 《Archives of microbiology》1989,152(3):280-288
The enzymology of methanol utilization in thermotolerant methylotrophic Bacillus strains was investigated. In all strains an immunologically related NAD-dependent methanol dehydrogenase was involved in the initial oxidation of methanol. In cells of Bacillus sp. C1 grown under methanol-limiting conditions this enzyme constituted a high percentage of total soluble protein. The methanol dehydrogenase from this organism was purified to homogeneity and characterized. In cell-free extracts the enzyme displayed biphasic kinetics towards methanol, with apparent K
m values of 3.8 and 166 mM. Carbon assimilation was by way of the fructose-1,6-bisphosphate aldolase cleavage and transketolase/transaldolase rearrangement variant of the RuMP cycle of formaldehyde fixation. The key enzymes of the RuMP cycle, hexulose-6-phosphate synthase (HPS) and hexulose-6-phosphate isomerase (HPI), were present at very high levels of activity. Failure of whole cells to oxidize formate, and the absence of formaldehyde-and formate dehydrogenases indicated the operation of a non-linear oxidation sequence for formaldehyde via HPS. A comparison of the levels of methanol dehydrogenase and HPS in cells of Bacillus sp. C1 grown on methanol and glucose suggested that the synthesis of these enzymes is not under coordinate control.Abbreviations RuMP
ribulose monophosphate
- HPS
hexulose-6-phosphate synthase
- HPI
hexulose-6-phosphate isomerase
- MDH
methanol dehydrogenase
- ADH
acohol dehydrogenase
- PQQ
pyrroloquinoline, quinone
- DTT
dithiothreitol
- NBT
nitrobluetetrazolium
- PMS
phenazine methosulphate
- DCPIP
dichlorophenol indophenol 相似文献
6.
Glutathione reductase fromSaccharomyces cerevisiae undergoes redox interconversionin situ andin vivo
José Peinado Javier Florindo J. López-Barea 《Molecular and cellular biochemistry》1992,110(2):135-143
Redox interconversion of glutathione reductase was studiedin situ withS. cerevisiae. The enzyme was more sensitive to redox inactivation in 24 hour-starved cells than in freshly-grown ones. While 5 μM NADPH
or 100 μM NADH caused 50% inactivation in normal cells in 30 min, 0.75 μM NADPH or 50 μM NADH promoted a similar effect in
starved cells. GSSG reactivated the enzyme previously inactivated by NADPH, ascertaining that the enzyme was subjected to
redox interconversion. Low EDTA concentrations fully protected the enzyme from NADPH inactivation, thus confirming the participation
of metals in such a process. Extensive inactivation was obtained in permeabilized cells incubated with glucose-6-phosphate
or 6-phosphogluconate, in agreement with the very high specific activities of the corresponding dehydrogenases. Some inactivation
was also observed with malate, L-lactate, gluconate or isocitrate in the presence of low NADP+ concentrations.
The inactivation of yeast glutathione reductase has also been studiedin vivo. The activity decreased to 75% after 2 hours of growth with glucono-δ-lactone as carbon source, while NADPH rose to 144%
and NADP+ fell to 86% of their initial values. Greater changes were observed in the presence of 1.5 μM rotenone: enzymatic activity
descended to 23% of the control value, while the NADH/NAD+ and NADPH/NADP+ ratios rose to 171% and 262% of their initial values, respectively. Such results indicate that the lowered redox potential
of the pyridine nucleotide pool existing when glucono-δ-lactone is oxidized promotesin vivo inactivation of glutathione reductase. 相似文献
7.
R. M. Alvarez B. Rodríguez J. M. Romano A. O. Díaz E. Gómez D. Miró L. Navarro G. Saura J. L. García 《World journal of microbiology & biotechnology》1992,8(2):214-215
Microbial lipids produced byRhodotorula glutinis grown in continuous culture with molasses under nitrogen-limiting conditions were evaluated and the effects of growth rate on fatty acid composition were studied. As the growth rate decreased, cell biomass, lipid content and lipid yield gradually increased. The maximum lipid content recorded was 39% (w/w) of dry cell biomass at a dilution rate of 0.04 h–1. The growth rate also affected fatty acid composition: oleic acid decreased with decreasing growth rate while stearic acid increased. 相似文献
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Mátyás Mink 《Biochemical genetics》1988,26(7-8):503-510
Physical mapping of the mitochondrial DNA of the wild-typeSaccharomyces cerevisiae strainRXII revealed that most of the restriction sites as well as the location of the apocytochromeb gene were identical in comparison with the known maps of the mitochondrial genome in otherSaccharomyces cerevisiae strains. In the middle of theSalI linearized map of theRXII mitochondrial DNA, a deletion was detected which resulted in the loss of twoEcoRI and oneBamHI restriction sites. The corresponding region, however, exists in most other laboratory strains ofSaccharomyces mapped so far. This region overlaps the introns aI2 and aI3 surrounding exon A3 sequences of the subunit 1 of the cytochrome
oxidase gene. The nucleotide sequence of the subunit 1 gene showed that theBamHI site was located close to the aI3-A4 intron-exon junction and the distalEcoRI site close to the aI2-A2 boundary. I therefore conclude that these two introns are deleted in the mitochondrial genome
of strainRXII. The exon A3 must have been conserved since this strain was respiratory competent. This result, while being a good example
of the morphological diversity of a genome with the same function, may contribute to an understanding of the role of introns
in the mitochondrial split genes in yeast. 相似文献
10.
S. G. Ball R. B. Wickner G. Cottarel M. Schaus C. Tirtiaux 《Molecular & general genetics : MGG》1986,205(2):326-330
Summary The chorismate mutase structural gene, ARO7, which is necessary for both phenylalanine and tyrosine biosynthesis was cloned by complementation in yeast. Genetic analysis showed that ARO7 was identical to a gene necessary for growth in hypertonic medium, OSM2, which mapped nearby. After restriction mapping and subcloning of the plasmid, the cloned gene was used to detect mRNA levels in several growth conditions. Enzyme activities were measured in various genotypes. At our level of detection ARO7-OSM2 is a low level constitutively expressed gene. 相似文献