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51.
52.
Effects of fructose on human fibroblast metabolism: the application of DNA measurements as a basis for interpretation 总被引:2,自引:0,他引:2
Robert Z. Eanes 《In vitro cellular & developmental biology. Plant》1985,21(6):328-332
Summary A fluorometric procedure for measuring DNA was used to study growth and metabolic responses of eight cell strains of human
foreskin fibroblasts. In preliminary studies this procedure gave more precise specific activity changes inN-acetyl-β-d-glucosaminidase (NAG) than did a protein activity basis, when changes in this enzyme's specific activity were investigated
as a function of experimental cell manipulation. When fibroblast growth in eight cell strains was compared in 134 mM
d-fructose vs. 13.4 mM glucose-supplemented minimum essential media, a significant increase in cellular DNA (50%) and protein (45%) occurred over
an 11-d period. No significant differences in media pH change, lactate production, or carbohydrate uptake occurred on a DNA
basis when cell metabolism was compared over the last 24 h of culture in the two media. Cells grown in fructose-containing
media tended to show a reduction in NAG specific activity when compared with those grown in glucose-containing media. 相似文献
53.
Summary Scanning cytophotometry following Feulgen-staining was used to determine nuclear DNA content in many differentiated tissues of nine cultivars, hybrids or selfed lines ofHelianthus annuus. Apart from such ephemeral tissues as endosperm and anther tapetum, it was found that tissue differentiation in sunflower occurs in the diploid condition, cells being arrested in the DNA presynthetic phase (G1). In certain cases, however, the nuclear DNA content of differentiated G1 cells does not exactly match the 2C DNA content found in meristematic cells, but may be either higher or lower. In endosperm and anther tapetum cells, nuclear DNA content may be as high as 24 C and 32 C, respectively. Cytological and autoradiographic analyses after3H-thymidine incorporation reveal that polyploidy in the tapetal cells is due to chromosome endoreduplication. No detectable difference between male-fertile and male-sterile plants exists as far as occurrence and level of cell polyploidy are concerned. The results are discussed in the context of previous investigations on the nuclear condition of differentiatedHelianthus annuus tissue. 相似文献
54.
Peter Nichols Benne K. Stulp J. Gwynfryn Jones David C. White 《Archives of microbiology》1986,146(1):1-6
DNA hybridization experiments showed that there was a high degree of homology amongVitreoscilla strains but not with DNA fromFilibacter limicola. Flexibacter spp were much more heterogeneous indicating a low genetic similarity. These results were also reflected in the membrane fatty acids of the bacteria. TheVitreoscilla strains were very similar with the 16:17c fatty acid being dominant. The membrane fatty acids ofF. limicola were dominated by a15:0 and a17:0 components which provided additional support for its relatedness to the genusBacillus. There was much greater diversity in the fatty acid patterns of theFlexibacter spp.F. aurantiacus, F. ruber andF. elegans shared the common dominant fatty acids 16:17c with theVitreoscilla strains, but this was replaced by the 16:16c acid inF. flexilis. F. ruber was distinguished by the absence of branched odd-chain monounsaturated fatty acids andF. elegans by the dominance of the -OH i15:0 acid. Precise determination of fatty acid double bond positions and geometry are essential for correct interpretation of increasingly complex ecological and taxonomic data sets. 相似文献
55.
Isolation and characterization of a human interleukin 2 gene 总被引:1,自引:0,他引:1
S Mita S Maeda K Obaru N Nishino K Shimada T Hirano K Onoue T Ogawa H Ogawa 《Biochemical and biophysical research communications》1983,117(1):114-121
An interleukin 2 (IL-2) gene was isolated from a Charon 4A human gene library. Electron microscopic examination of 15 heteroduplexes formed between the genomic DNAs and the IL-2 cDNAs demonstrated that the size of the IL-2 gene is about 5.1 +/- 0.5 kb and that there are at least two introns in this gene. Nucleotide sequence of the 5' flanking region of the IL-2 gene showed a homology with that of the corresponding region of the human immune interferon gene. 相似文献
56.
Anna Maria Tagliasacchi Laura Maria Costantina Forino Manuela Frediani Silvana Avanzi 《Protoplasma》1983,115(2-3):95-103
Summary The pattern of DNA and RNA puffs in pair VII of polytene chromosomes has been investigated in the suspensor ofPhaseolus coccineus during early embryo development. The pattern of3H-TdR and3H-U incorporation has been also detected. Collected data indicate that: 1. both heterochromatic regions, p11 and q(111+112), of chromosome pair VII, organize large DNA puffs; 2. DNA puffs of both regions are specific of different embryo differentiation steps; 3. a seasonal influence on the DNA puffing seems also to be present, as demonstrated by the comparison of the results collected in two different crops; 4. the incorporation experiment by3H-TdR evidences that not all DNA puffs show clustered labeling; 5. the RNA puffing of the two regions seems also to be specific of determined embryo stages. 相似文献
57.
A simple method for electrophoretic elution of nucleic acids from gel slices is described. The procedure utilizes a standard tube gel system and can be completed in as little as one hour. Nucleic acids are recovered in a small volume with almost 100% efficiency. The procedure is applicable equally to acrylamide and agarose gels, and small as well as large RNA and DNA molecules. The eluted nucleic acids are essentially undegraded and are suitable for a variety of structural and biological analyses. 相似文献
58.
G. Stauder H. Riesemann W.M. Joester K.E. Joester 《Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression》1983,741(3)
A third DNA polymerase ‘C’ with low molecular weight was isolated and purified 3700-fold from ground hyphae of Neurospora crassa WT 74 A, which shows similarities to β- and γ-polymerases from higher eukaryotes: preference for poly(rA)(dT) as a template/primer, inhibition by p-chloromercuribenzoate, resistance against N-ethylmaleimide up to 10 mmol/l, and molecular weight of about 40 000. This polymerase elutes as a distinct peak from DEAE-cellulose at 0.60 mol/l KCl and has an optimum for K+ at 2–20 mmol/l, for Mn2+ at 0.8 mmol/l, for Mg2+ at 4.0 mmol/l, the pH optimum is 8.0. Its Km is 1.5 μmol/l using dTTP as substrate. The enzyme activity described here is free of endonuclease but contains detectable amounts of exonuclease. 相似文献
59.
RICHARD A. ALBACH 《The Journal of eukaryotic microbiology》1989,36(2):197-205
This review will concentrate on certain aspects of the nucleic acids of Entamoeba histolytica. Utilization and synthesis of purines and pyrimidines will initially be briefly discussed, e.g. salvage vs. de novo pathways, uptake studies and recognition of at least 4 transport loci. Data will be presented which show that the distribution and synthesis of RNA (to a lesser extent DNA) in the nucleus is basically the opposite one finds in other eukaryotes, viz. most RNA (ribosomal?) is synthesized (or accumulates) in the peripheral chromatin (functional equivalent of nucleolus?). The DNA is distributed and synthesized primarily throughout the nucleus. It is usually so dispersed that it will not stain with e.g. the standard Feulgen technique, unless the DNA condenses around the endosome (not a nucleolar equivalent) prior to nuclear division. Isolation of rRNA was difficult due, in part, to potent and difficult to inhibit RNase(s), some of which are apparently intimately bound to ribosomal subunits. The 25S (1.3 kDa), 178 (0.8 kDa) and 58 rRNA were recovered after isolation with a high salt SDS-DEP technique. This is the only procedure which enables us to obtain high yields of 258 rRNA: guanidine or guanidinium which permits isolation of intact functional mRNA results in isolation of small amounts of 28 RNA relative to 178 RNA. The 258 RNA is “nicked” (apparently during nuclear processing) and dissociates readily into 1 78 (0.7 kDa) and 168 (0.6 kDa) species, and a more rigidly bound 5.88 species. A small amount of “unnicked” 258 RNA was recovered with guanidine. Two DNA-dcpendent RNA polymerases (I and II) with a pronounced preference for denatured DNA as template were eluted from DEAE-Sephadex in reverse order of what occurs in other eukaryotes, except Physarum polycephalum. This conclusion was based on salt optima and alpha-amanitin sensitivity studies. Initial characterization of DNA isolated with a procedure capable of isolating > 100-kbp Leishmania DNA showed that undigested DNA migrates as a broad band between markers 6 and 24 kbp. The persistent recovery of such a “band” by us and Perez-Mutul et al. no larger than ca. 24 kbp (with the exception of >48 kbp DNA isolated by Hernandez et al. using an in situ lysis technique which did not include a proteinase) may be due to nicks introduced during isolation; or, perhaps much of the amebal DNA exists in vivo as gene sized fragments. However, preliminary data generated using orthogonal pulse-field agarose gel electrophoresis do suggest that amebal DNA may be in small chromosomes. 相似文献
60.
Tsuneyoshi Kuroiwa 《Journal of plant research》1989,102(2):291-329
It has been established that organelles, such as mitochondria and plastids, contain organelle-specific DNA and arise from
the division of pre-existing organelles (e.g., Possingham and Lawrence, 1983). We propose that organelle DNAs, such as mitochondrial
DNA and plastid DNA are not naked in organellesin situ but are organized in each case to form an “organelle nucleus” with basic proteins (Kuroiwa, 1982). The concept of organelle
nuclei has changed our ideas about the division of organelles. Thus, the process of organelle division must be composed of
two main events: division of the organelle nucleus and organellekinesis (division of the other components of the mitochondrion
or plastid). The latter term has been adopted as an appropriate analogue of cytokinesis.
We were the first to identify the plastid-dividing ring (PD-ring), which is located in the cytoplasm close to the outer envelope
membrane at the constricted isthmus of dividing chloroplasts in the red algaCyanidium caldirum. The PD-ring is about 60 nm in width and 25 nm in thickness, and is a circular bundle of actin-like, fine filaments, each
about 4–5 nm in diameter. Since cytochalasin B, an inhibitor of polymerization of actin filaments, inhibits the formation
of the PD-ring and, thus, prevents subsequent division of chloroplasts, the PD-ring is thought to be a structure that is essential
for the division of plastids (plastidkinesis).
The behavior of the PD-ring during a cycle of chloroplast division can be classified into the following four stages on the
basis of morphological and temporal differences. The chloroplast growth stage: the small, spherical chloroplast increases
in volume and becomes a football-like structure, while the PD-ring from the previous division disappears. Formation of the
PD-ring: the somewhat electron-dense body (see below) is fragmented into many, somewhat electron-dense granules, which are
aligned along the equatorial region of the chloroplast and fine filaments are formed from the somewhat electron-dense granules
in the equatorial region. The fine filaments of the PD-ring align themselves according to the longest axis of their overall
domain, i.e., circumferentially. Contraction stage: a bundle of fine filaments begins to contract and generates a deep furrow.
Conversion stage: after chloroplast division, the remnants of the PD-ring are converted into somewhat electron-dense bodies.
Similar events occur during the second cycle of chloroplast division. Since similar structures are observed extensively in
the plastids of algae, moss and higher plants, the PD-ring appears to be an essential structure for the division of plastids
in plants. 相似文献