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51.
Klaus Werner Wolf Sabine Glatzel Frauke Niedereichholz Bryan M. Turner 《Biotechnic & histochemistry》1996,71(3):137-144
We describe a method for isolating chromosomes from testes of the desert locust, Schistocerca gregaria, and their subsequent incubation with antibodies directed against chromosomal proteins. The procedure involves hypotonic pretreatment of the germ cells, centrifugation onto coverslips in a cytocentrifuge and immunolabeling, while still unfixed, using a chromatin-stabilizing buffer. In the present case, an antibody specific for the acetylated isoforms of his tone H4 was tested. After the antibody treatment, the preparations are fixed using formaldehyde, stained with a DNA-specific fluorescent dye and mounted. Analysis of the preparations revealed good preservation of chromosome structure in prophase spermatogonia and late prophase I spermatocytes. Fully condensed chromosomes were not observed and are probably lost during preparation. The bright fluorescence of the autosomes indicates that the reaction between the antibody against acetylated histone H4 and its chromosomal antigen is not impeded. In contrast, the X univalent remained unstained with the exception of a small terminal band. Thus, cytospin preparations of locust germ cells allow high resolution immunolabeling with antibodies against chromosome-associated proteins. 相似文献
52.
A new method was developed to purify the streptococcal hyaluronate synthase in active form to electrophoretic homogeneity. The method is based on the extraction of protoplast membranes with digitonin and a phase separation into an aqueous and a detergent phase induced by addition of polyethylene glycol 6000 at 0°C. Proteins bound to hyaluronate were enriched in the aqueous phase, whereas other membrane proteins resided in the detergent phase. Final purification of the hyaluronate synthase was achieved by ion exchange chromatography. 相似文献
53.
Sabine J. Rundle Andrew J. Hartung John W. Corum III Michael O'Neill 《Plant molecular biology》1995,28(2):257-266
Type 2A serine/threonine protein phosphatases (PP2A) are key components in the regulation of signal transduction and control of cell metabolism. The activity of these protein phosphatases is modulated by regulatory subunits. While PP2A activity has been characterized in plants, little is known about its regulation. We used the polymerase chain reaction to amplify a segment of a cDNA encoding the B regulatory subunit of PP2A from Arabidopsis. The amplified DNA fragment of 372 nucleotides was used as a probe to screen an Arabidopsis cDNA library and a full-length clone (AtB) of 2.1 kbp was isolated. The predicted protein encoded by AtB is 43 to 46% identical and 53 to 56% similar to its yeast and mammalian counterparts, and contains three unique regions of amino acid insertions not present in the animal B regulatory subunit. Genomic Southern blots indicate the Arabidopsis genome contains at least two genes encoding the B regulatory subunit. In addition, other plant species also contain DNA sequences homologous to the B regulatory subunit, indicating that regulation of PP2A activity by the 55 kDa B regulatory subunit is probably ubiquitous in plants. Northern blots indicate the AtB mRNA accumulates in all Arabidopsis tissues examined, suggesting the protein product of the AtB gene performs a basic housekeeping function in plant cells. 相似文献
54.
In order to better understand the biochemical mechanisms of DNA metabolism in chloroplasts, repair of UV induced plastome damage in vivo was determined by exposure of soybean suspension cells to UV light and subsequent quantitation of the damage remaining in nuclear and chloroplast encoded genes with time by quantitative polymerase chain reaction (QPCR). The kinetics of damage rapir in the nuclear rbcS gene suggest that photoreactivation and dark mechanisms are active, while for the plastome encoded psbA gene only a light-dependent repair process was detected which is considerably slower than would be expected for photolyase-mediated photoreactivation. 相似文献
55.
Sabine Lhernould Yannis Karamanos Patrice Lerouge Henri Morvan 《Glycoconjugate journal》1995,12(1):94-98
The peptide-N
4-(N-acetylglucosaminyl) asparagine amidase (PNGase Se) earlier described [Lhernould S., Karamanos Y., Bourgerie S., Strecker G., Julien R., Morvan H. (1992)Glycoconjugate J
9:191–97] was partially purified from culturedSilene alba cells using affinity chromatography. The enzyme is active between pH 3.0 and 6.5, and is stable in the presence of moderate concentrations of several other protein unfolding chemicals, but is readily inactivated by SDS. Although the enzyme cleaves the carbohydrate from a variety of animal and plant glycopeptides, it does not hydrolyse the carbohydrate from most of the corresponding unfolded glycoproteins in otherwise comparable conditions. The substrate specificity of this plant PNGase supports the hypothesis that this enzyme could be at the origin of the production of unconjugated N-glycans in a suspension medium of culturedSilene alba cells.Abbreviations GlcNAc
N-acetylglucosamine
- PNGase
peptide-N
4-(N-acetylglucosaminyl) asparagine amidase
- BSA
bovine serum albumin
- SDS-PAGE
sodium dodecyl sulfate-polyacrylamide gel electrophoresis
- TLC
thin layer chromatography
- HPAEC-PAD
High pH anion exchange chromatography-pulsed amperometric detection 相似文献
56.
Sabine Castano Bernard Desbat Isabelle Cornut Philippe Méléard Jean Dufourcq 《Letters in Peptide Science》1997,4(4-6):195-200
De novo designed extremely simplified amphipathic basicLeuiLysj (i = 2j) peptides of 8, 9 and 15residues were synthesized to clarify the mechanism of action of naturalcytotoxic and hemolytic small proteins or peptides. They proved to havestrong hemolytic activity towards human erythrocytes which increases withpeptide length. These peptides are highly surface active and form stablepeptidic films at the air/water interface. The sensitive and efficient FTIRmodulated polarization technique (PMIRRAS) allows one to obtain in situstructural and orientational information about the peptides at theinterface. A transition of secondary structure is observed: the shorterpeptides (8 and 9 residues) adopt -sheet structures while the longerone (15 residues) is folded into an -helix. In both cases, the peptideslie with the axis parallel to the interface. Their insertion into adimyristoylphosphatidylcholine monolayer can be followed from the increasein the surface and/or pressure of the films. In the mixed films, thepeptides adopt the same structure and orientation as observed at theair/water interface. Therefore, among the same series of peptides, atransition from -sheet to -helix occurs when the length increases(roughly >10 aa), but despite this drastic change both types ofstructures result in strongly hemolytic peptides. 相似文献
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60.
Sabine Karg 《Vegetation History and Archaeobotany》1995,4(1):41-50
Weed assemblages from late medieval cornfields have been studied for the first time in northern Switzerland. Eleven samples from at least two different grain stores were investigated. The samples were collected from the carbonised remains of six wooden houses built in the late 13th century A.D. and which burnt down in the middle of the 15th century. The weed floras found in the spelt (Triticum spelta) and oats (Avena sativa) indicate a high botanical diversity in the cornfields at harvest time. Although oats are normally a summer crop and spelt a winter crop, both summer and winter crop weeds (as well as many different present-day grassland taxa) were found in each type of grain. Most of the weeds found were perennial plants, which was interpreted as an indication of both extensive tillage of the arable land and application of the three-field rotation system (Dreifelderwirtschaft). The spectra of the two palaeophytocoenoses (assemblages of ancient plant remains) studied suggest that the phytosociological method may not be reliable for classification of the late medieval remains into summer and winter crop weed communities. These findings should provide a better understanding of the development of anthropogenic plant communities, and in particular, the development of crop weed communities. 相似文献