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21.
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The polymerase chain reaction was used to amplify the staphylococcal enterotoxin B and C genes (entB and entC1) and the staphylococcal nuclease gene (nuc). Two sets of primers ("nested primers") were found to be necessary for the detection of low copy numbers of purified DNA in diluent. These allowed detection of ca. 1 fg of purified target DNA, while 100 pg was required before detection of entB, entC1, and nuc with single primer pairs was possible. With nested primers, enterotoxigenic Staphylococcus aureus cells could be detected in artificially contaminated dried skimmed milk samples at levels of ca. 10(5) CFU ml-1 within 8 h. No cross-reaction was observed between the highly homologous entB and entC1 genes. The method showed total specificity for entC1 when tested against a wide variety of other bacteria. 相似文献
23.
Cold acclimation and cold-regulated gene expression in ABA mutants of Arabidopsis thaliana 总被引:22,自引:0,他引:22
We have examined the cold-induced enhancement of freezing tolerance and expression of cold-regulated (cor) genes in Arabidopsis thaliana (L.) Heynh (Landsberg erecta) and abscisic acid (ABA)-deficient (aba) and ABA-insensitive (abi) mutants derived from it. The results indicate that the abi mutations had no apparent effect on freezing tolerance, while the aba mutations did: cold-acclimated aba mutants were markedly impaired in freezing tolerance compared to wild-type plants. In addition, it was observed that non-frozen leaves from both control and cold-treated aba mutant plants were more ion-leaky than those from corresponding wild-type plants. These data are consistent with previous observations indicating that ABA levels can affect freezing tolerance. Whether ABA has a direct role in the enhancement of freezing tolerance that occurs during cold acclimation, however, is uncertain. Several studies have suggested that ABA might mediate certain changes in gene expression that occur during cold acclimation. Our data indicate that the ABA-induced expression of three ABA-regulated Arabidopsis cor genes was unaffected in the abi2, abi3, and aba-1 mutants, but was dramatically impaired in the abi1 mutant. Cold-regulated expression of all three cor genes, however, was nearly the same in wild-type and abi1 mutant plants. These data suggest that the cold-regulated and ABA-regulated expression of the three cor genes may be mediated through independent control mechanisms. 相似文献
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25.
In this paper we examined the effects of transforming growth factor β (TGFβ) on the proliferation and differentiation of rabbit tracheal epithelial cells in primary culture. Treatment of these cells with TGFβ inhibits cell proliferation in a time- and dose-dependent manner; concentrations as low as 1 pM are able to inhibit cell growth. Concomitantly, TGFβ causes cells to accumulate in the G0/G1 phase of the cell cycle and a sharp reduction in the ability of the cells to form colonies after subculture at clonal density. These results indicate that TGFβ induces terminal cell division in these cells. The inhibition of cell growth is accompanied by changes in cell morphology and a stimulation of the formation of cross-linked envelopes. TGFβ enhances the levels of transglutaminase activity and cholesterol sulfate, two markers of squamous differentiation. Our results indicate that TGFβ induces terminal squamous cell differentiation in rabbit tracheal epithelial cells. Retinoic acid (RA) does not affect the commitment to terminal cell division induced by TGFβ, but inhibits the expression of the squamous phenotype. Growth of normal human bronchial epithelial cells was affected by TGFβ in a way similar to that of rabbit tracheal epithelial cells. Several carcinoma cell lines tested were quite resistant to TGFβ, whereas growth of one carcinoma cell line was stimulated by TGFβ. These results indicate that a modified response to TGFβ could be one mechanism involved in the aberrant growth control of malignant cells. 相似文献
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27.
Gibberellin metabolism in cell-free extracts from spinach leaves in relation to photoperiod 总被引:19,自引:12,他引:7
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Cell-free extracts capable of converting [14C]-labeled gibberellins (GAs) were prepared from spinach (Spinacia oleracea L.) leaves. [14C]-labeled GAs, prepared enzymically from [14C]mevalonic acid, were incubated with these extracts, and products were identified by gas chromatography-mass spectrometry. The following pathway was found to operate in extracts from spinach leaves grown under long day (LD) conditions: GA12 → GA53 → GA44 → GA19 → GA20. The pH optima for the enzymic conversions of [14C]GA53, [14C]GA44 and [14C]GA19 were approximately 7.0, 8.0, and 6.5, respectively. These three enzyme activities required Fe2+, α-ketoglutarate and O2 for activity, and ascorbate stimulated the conversion of [14C]GA53 and [14C]GA19. Extracts from plants given LD or short days (SD) were examined, and enzymic activities were measured as a function of exposure to LD, as well as to darkness following 8 LD. The results indicate that the activities of the enzymes oxidizing GA53 and GA19 are increased in LD and decreased in SD or darkness, but that the enzyme activity oxidizing GA44 remains high irrespective of light or dark treatment. This photoperiodic control of enzyme activity is not due to the presence of an inhibitor in plants grown in SD. These observations offer an explanation for the higher GA20 content of spinach plants in LD than in SD. 相似文献
28.
Gibberellins (GAs) A17, A19, A20, A29, A44, 2OH-GA44 (tentative) and GA29-catabolite were identified in 21-day-old seeds of Pisum sativum cv. Alaska (tall). These GAs are qualitatively similar to those in the dwarf cultivar Progress No. 9 with the exception of GA19 which does not accumulate in Progress seeds. There was no evidence for the presence of 3-hydroxylated GAs in 21 day-old Alaska seeds. Dark-grown shoots of the cultivar Alaska contein GA1, GA8, GA20, GA29, GA8-catabolite and GA29-catabolite. Dark-grown shoots of the cultivar Progress No.9 contain GA8, GA20, GA29 and GA29-catabolite, and the presence of GA1 was strongly indicated. Quantitation using GAs labelled with stable isotope showed the level of GA1 in dark-grown shoots of the two cultivars to be almost identical, whilst the levels of GA20, GA29 and GA29-catabolite were significantly lower in Alaska than in Progress No. 9. The levels of these GAs in dark-grown shoots were 102- to 103-fold less than the levels in developing seeds. The 2-epimer of GA29 is present in dark-grown-shoot extracts of both cultivars and is not thought to be an artefact.Abbreviations cv
cultivar
- GAn
gibberellin An
- GC
gas chromatography
- GC-MS
combined gas chromatographymass spectrometry
- HPLC
high-pressure liquid chromatography
- KRI
Kovats retention index
- MeTMSi
methyl ester trimethylsilyl ether 相似文献
29.
30.
Jill C. Tarter 《Origins of life and evolution of the biosphere》1986,16(3-4):399-399