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21.
22.
Aminoglycoside suppression at UAG, UAA and UGA codons in Escherichia coli and human tissue culture cells 总被引:6,自引:0,他引:6
Robin Martin Anne E. Mogg Louise A. Heywood Lars Nitschke Julian F. Burke 《Molecular & general genetics : MGG》1989,217(2-3):411-418
Summary We have compared the suppression of nonsense mutations by aminoglycoside antibiotics inEscherichia coli and in human 293 cells. Six nonsense alleles of the chloramphenicol acetyl transferase (cat) gene, in the vector pRSVcat, were suppressed by growth in G418 and paromomycin. Readthrough at UAG, UAA and UGA codons was monitored with enzyme assays
for chloramphenicol acetyl transferase (CAT), in stably transformed bacteria and during transient expression from the same
plasmid in human 293 tissue culture cells. We have found significant differences in the degree of suppression amongst three
UAG codons and two UAA codons in different mRNA contexts. However, the pattern of these effects are not the same in the two
organisms. Our data suggest that context effects of nonsense suppression may operate under different rules inE. coli and human cells. 相似文献
23.
Anin situ method, derived from anin vivo method, was used to determine nitrate reductase activity (NRA) in:i) excised barley and corn shoots and excised soybean leaves
during a N-depletion experiment and; ii) roots and shoots of N-depleted barley and corn seedlings during induction of nitrate,
reductase (NR). Nitrate reduction, calculated from thesein situ RNA measurements, was compared with estimates of each organ's nitrate reduction in light aerobic conditions from NO
3
−
consumption and a15N model (Gojonet al., 1986b).
Thein situ RNA of roots strongly underestimated their15NO
3
−
reduction. In contrast, in barley and corn shoots and in the first trifoliolate leaves from 26-day-old, soybean, thein situ NRA assay gave a fair approximation of the true NO
3
−
reduction rate (relative differences ranging from −14 to +32%). In young soybean leaves (from 20-day-old plants), however,
thein situ NRA strongly underestimated the actual NO
3
−
reduction. The physiological significance of thein situ NRA assay in shoots and roots, and its value for field studies are discussed from these results. 相似文献
24.
Komandoor E. Achyuthan Ann Mary Robin Bhaermani Charles S. Greenberg 《Molecular and cellular biochemistry》1989,85(1):57-65
Calcium ions are crucial for expression of transglutaminase activity. Although lanthanides have been reported to substitute for calcium in a variety of protein functions, they did not replace the calcium requirement during transglutaminase activity measurements. Furthermore, lanthanides strongly inhibited purified liver transglutaminase activity using either casein or fibrinogen as substrates. Terbium (III) inhibition of transglutaminase-catalyzed putrescine incorporation into casein was not reversed by the presence of 10–200 fold molar excess of calcium ions (Ki for Tb(III)=60 µM). Conformational changes in purified liver transglutaminase upon Tb(III) binding were evident from a biphasic effect of Tb(III) on transglutaminase binding to fibrin. Low concentrations of Tb(III) (1 µM to 10 µM inhibited the binding of transglutaminase to fibrin, whereas higher concentrations (20 µM to 100 µM promoted binding. Conformational changes in purified liver transglutaminase consequent to Tb(III) binding were also demonstrated by fluorescence spectroscopy due to Forster energy transfer. Fluorescence emission was stable to the presence of 200 mM NaCl and 100 mM CaCl2 only partially quenched emission. Purified liver transglutaminase strongly bound to Tb(III)-Chelating Sepharose beads and binding could not be disrupted by 100 mM CaCl2 solution. Our data suggest that Tb(III)-induced conformational changes in transglutaminase are responsible for the observed effects on enzyme structure and function. The potential applications of Tb(III)-transglutaminase interactions in elucidating the structure-function relationships of liver transglutaminase are discussed. 相似文献
25.
A data base was compiled containing the amino acid sequences of 12 aspartate aminotransferases and 11 other aminotransferases. A comparison of these sequences by a standard alignment method confirmed the previously reported homology of all aspartate aminotransferases and Escherichia coli tyrosine aminotransferase. However, no significant similarity between these proteins and any of the other aminotransferases was detected. A more rigorous analysis, focusing on short sequence segments rather than the total polypeptide chain, revealed that rat tyrosine aminotransferase and Saccharomyces cerevisiae and Escherichia coli histidinol-phosphate aminotransferase share several homologous sequence segments with aspartate aminotransferases. For comparison of the complete sequences, a multiple sequence editor was developed to display the whole set of amino acid sequences in parallel on a single work-sheet. The editor allows gaps in individual sequences or a set of sequences to be introduced and thus facilitates their parallel analysis and alignment. Several clusters of invariant residues at corresponding positions in the amino acid sequences became evident, clearly establishing that the cytosolic and the mitochondrial isoenzyme of vertebrate aspartate aminotransferase, E. coli aspartate aminotransferase, rat and E. coli tyrosine aminotransferase, and S. cerevisiae and E. coli histidinol-phosphate aminotransferase are homologous proteins. Only 12 amino acid residues out of a total of about 400 proved to be invariant in all sequences compared; they are either involved in the binding of pyridoxal 5'-phosphate and the substrate, or appear to be essential for the conformation of the enzymes. 相似文献
26.
Pedro J. N. Silva Richard K. Koehn Walter J. Diehl III Robin P. Ertl Elaine B. Winshell Mauro Santos 《Biochemical genetics》1989,27(7-8):451-467
Four samples of the musselMytilus edulis were taken between 1984 and 1987 from Stony Brook, New York, and used to study the glucose-6-phosphate isomerase (GPI) polymorphism
in this species.In vitro specific activity andin vivo flux measured in the same animals were found to be significantly correlated. A significant effect of GPI genotype on flux
was observed in one of the samples; overall, significant evidence of effect of genotype on enzyme activity was also obtained.
GPI activities of common genotypes tend to deviate less from the population mean than those of rare (frequency less than 5%)
genotypes. This suggests the possibility that rare GPI genotypes are rare as a consequence of having biochemical properties
that deviate from an optimum level and, therefore, having a lower fitness. In support of this hypothesis, we found in one
of our samples that shell length is a concave function of GPI activity with an intermediate optimum activity level.
The financial support provided to P.J.N.S. by the Luso-American Educational Commission (Fulbright Program), the Instituto
Nacional de Investigacao Científica (Portugal), and the Faculdade de Ciências da Universidade de Lisboa during several stages
of this research is gratefully acknowledged. Financial support from the Ministerio de Educatión y Ciencia (Spain) in the form
of a postdoctoral Fulbright/MEC fellowship to M.S. is also gratefully acknowledged. Research was supported by National Science
Foundation Grant BSR-8415060 to R.K.K. This is contribution No. 736 from the Program in Ecology and Evolution, State University
of New York at Stony Brook.
On leave from Departamento de Biologia Vegetal, Faculdade de Ciências, Universidade de Lisboa, Campo Grande C2, Lisboa, Portugal. 相似文献
27.
Menstrual-cycle phase and sexual behavior in semi-free-ranging stumptail macaques (Macaca arctoides)
The sexual behavior and female reproductive cycles of a group of island-dwelling stumptail macaques (Macaca arctoides)were monitored over a 6-month period, yielding 530 observation hr and 268 copulations. Compared to nondominant males, the
dominant male copulated at a relatively high rate throughout the cycle, but largely with one high-ranking female. The non-dominant
males copulated most frequently at midcycle. Female presenting was highest at midcycle, but only to the dominant male. Cross-study
discrepancies may be due to different observation methods and restricted environmental conditions that mask female-initiated
sexual behavior. The more naturalistic setting of this study allowed for a fuller expression of proceptivity. Contrary to
some previous conclusions, present findings suggest that both hormonal and socioenvironmental factors influence the patterns
of sexual behavior found in stumptail macaque colonies. 相似文献
28.
29.
Jeanine Brederoo Piet de Wildt Corrie Popp-Snijders Robin F. Irvine Alan Musgrave Herman van den Ende 《Planta》1991,184(2):175-181
InChlamydomonas eugametos gametes, phosphatidylinositol 4-phosphate (PtdInsP) and phosphatidylinositol 4,5-bisphosphate (PtdInsP2) comprised 0.4 and 0.3% of the whole-cell phospholipids. They were concentrated in the plasma membrane around the cell body and were present in low concentrations in the flagellar membrane. When gametes were fed32PO
4
-
, the label was rapidly incorporated into PtdInsP and PtdInsP2 and only slowly incorporated into structural lipids such as phosphatidylethanolamine and phosphatidylglycerol. Similarly, when a pulse of32PO
4
-
was chased with PO
4
-
, the label was rapidly lost from the polyphosphoinositol lipids but not from the structural lipids. The major fatty acids in the polyphosphoinositides were C-22 carbon polyenoic acids (70%). The significance of these results in relationship to intracellular signalling via inositol phosphates and Ca2+ is discussed.Abbreviations InsP3
inositol 1,4,5-trisphosphate
- mt–/mt+
mating-type plus or minus
- PtdA
phosphatidic acid
- PtdEtn
phosphatidylethanolamine
- PtdGro
phosphatidylglycerol
- PtdIns
phosphatidylinositol
- PtdInsP
phosphatidylinositol 4-phosphate;
- PtdInsP2
phosphatidylinositol 4,5-bisphosphate
- TCA
trichloroacetic acid
We thank Frank Schuring for Fig. 5A and Susan Kenter, Hans Kruisselbrink, Saskia Bijvank and Nelleke Corbett for their enthousiastic assistance. 相似文献
30.
After total-body exposure to various doses of ionizing radiation, the ability of lymphocytes to interact specifically with high endothelial venules of rat cervical and mesenteric lymph nodes was analyzed in frozen sections. Following a radiation dose of 1.5 Gy, high endothelial venules remained intact and the binding of unirradiated lymphocytes to the venules was enhanced relative to unirradiated controls. At radiation doses above 5.0 Gy, damage to high endothelial venules was observed histologically as well as assessed functionally. There was a significant decrease in specific lymphocyte-venule binding and a significant increase in nonspecific binding. These findings suggest that radiation-induced damage to high endothelial venules might play a role in radiation-induced immunosuppression by interfering with the normal passage of lymphocytes from the blood into lymph nodes via a specific interaction between lymphocytes and high endothelial venules. 相似文献