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81.
Collection and quality of rhesus monkey semen 总被引:2,自引:0,他引:2
S E Lanzendorf P M Gliessman A E Archibong M Alexander D P Wolf 《Molecular reproduction and development》1990,25(1):61-66
Electroejaculation is an accepted method of semen collection from nonhuman primates. Although both penile and rectal probe stimulation techniques have been used, there has been a general lack of consistency and detail regarding their application. This report describes the collection, processing, and evaluation of rhesus monkey semen contrasting two methods of penile electroejaculation: 1) a constant-voltage method where stimulus current is a variable and 2) a constant-current method where stimulus current is operator-controlled. The constant-current method was the more efficient procedure, requiring a lower stimulus current for successful electroejaculation. The influence on semen quality of potentially toxic agents used in the procedure, surgical glove powder and electrolyte cream, was tested; both were detrimental as measured by motility loss. No correlation was found between coagula volume and sperm numbers. The intra- and interanimal variability in semen samples from six monkeys was also evaluated. Penile electroejaculation, combined with control of stimulus current, provides a consistent, successful, and humane method for the collection of semen in the rhesus monkey. 相似文献
82.
The role of phosphatases in signal transduction 总被引:10,自引:0,他引:10
D R Alexander 《The New biologist》1990,2(12):1049-1062
The importance of phosphatases in regulating the phosphorylation of proteins involved in cell signaling has been demonstrated by four recent discoveries. First, a new family of receptor-like transmembrane phosphotyrosine phosphatases, highly conserved throughout evolution, was shown to be distributed in a wide variety of tissues. Extensive heterogeneity in the extracellular regions of these molecules points to the existence of a wide diversity of ligands. These ligands are thought to mediate transduction of signals to the cell interior by means of the phosphatase activity occurring within the cytoplasmic domains of the receptor-like transmembrane phosphotyrosine phosphatases. Second, cell-permeable tumor promoters, such as okadaic acid, were shown to be potent phosphatase inhibitors that have multiple effects on signaling pathways. Third, the subunits of the type 2A phosphatase were found to associate with transforming antigens encoded by DNA tumor viruses, indicating a role for phosphatases in mediating abnormal proliferative events. Fourth, several cell-cycle mutants were found to encode phosphatases. This review focuses on the significance of the transmembrane phosphotyrosine phosphatases and on the possible ways in which intracellular phosphatases function in signaling pathways. 相似文献
83.
Gerlinde Linne von Berg Alexander Samoylov Manfred Klaas Peter Hanelt 《Plant Systematics and Evolution》1996,200(3-4):253-261
The utility of chloroplast DNA variation for checking a recently proposed infrageneric classification of the genusAllium was tested. cpDNA restriction patterns of 49 species representing the main subgenera, sections, and subsections of the existing classification were compared. 363 different fragments generated by 4 restriction enzymes were identified and analysed by UPGMA clustering. The resulting phenogram largely confirms the subgeneric classification based on an integration of morphological and other methods. 相似文献
84.
85.
Dorozynski A 《BMJ (Clinical research ed.)》1994,308(6940):1319-1320
86.
Dorozynski A 《BMJ (Clinical research ed.)》1994,308(6933):873-874
87.
Matthias Liebergesell Alexander Steinbüchel 《Applied microbiology and biotechnology》1993,38(4):493-501
From a genomic library of Thiocystis violaceae strain 2311 in L47, two adjacent EcoRI restriction fragments of 5361 base pairs (bp) and of 1978 bp were cloned. The 5361-bp EcoRI restriction fragment hybridized with a DNA fragment harbouring the Alcaligenes eutrophus poly(3-hydroxyalkanoate) (PHA) synthase operon (phbCAB) and restored the ability to synthesize and accumulate PHA in PHA-negative mutants derived from A. eutrophus. The nucleotide sequence analysis of both fragments revealed five open-reading frames (ORFs); at least three of them are probably relevant for PHA biosynthesis. The amino acid sequences of the putative proteins deduced from these genes indicate that they encode a -ketothiolase [phbA
Tv, relative molecular mass (Mr) 40850], which exhibited 87.3% amino acid identify with the -ketothiolase from Chromatium vinosum. The amino acid sequences of the putative proteins deduced from ORF2Tv (Mr 41 450) and phbC
Tv (Mr 39 550), which were located upstream of and antilinear to phbA
Tv, exhibited 74.7% and 87.6% amino acid identify, respectively, with the corresponding gene products of C. vinosum. Downstream of and antilinear to phbC
Tv was located ORF5, which encodes for a protein of high relative molecular mass (Mr 76428) of unknown function. With respect to the divergent organisation of ORF2Tv and phbC
Tv on one side and of phbA
Tv on the other side and from the homologies of the putative gene products, this region of the T. violaceae genome resembled very much the corresponding region of C. vinosum, which was identified recently.
Correspondence to: A. Steinbüchel 相似文献
88.
A cosmid gene bank of partially EcoRI-digested genomic DNA from Methylobacterium extorquens IBT no. 6 was screened for DNA fragments restoring polyhydroxyalkanoic-acid (PHA) accumulation in the PHA-negative mutant Alkaligenes eutrophus H16 PHB–4. The M. extorquens PHA-synthase structural gene phaC
Mex
was mapped on a 23-kbp EcoRI fragment by complementation studies, by hybridization experiments with heterologous DNA probes from A. eutrophus H16 encoding for phaA, phaB and phaC and by nucleic acid sequence analysis. Evidence for the presence of genes for a -ketothiolase or an acetoacetyl-coenzyme A reductase on this fragment was not obtained. The nucleotide sequence of a 3.7-kbp region was obtained. It contained the entire 1.815-kbp phaC
Mex plus approximately each 900-bp upstream and downstream of phaC
Mex. PhaC
Mex
encoded a protein of 605 amino acods with a relative molecular mass (Mr) of 66742, which exhibited 38.1% amino acid identity with the A. eutrophus PHA synthase. Determination of the N-terminal amino acid sequence of an Mr 65 000 protein, which was enriched concomitantly with the purification of PHA granules in sucrose gradients, revealed a sequence that was identical with the amino acid sequence deduced from the most probable translation start codon except for a valine, which was obviously removed post-translationally. Enzyme analysis, which was done with the native gene and a phaC
Mex
-lacZ fusion gene, gave no evidence for expression of phaC
Mex
in Escherichia coli. 相似文献
89.
Alexander A. DiIorio Pamela J. Weathers Ronald D. Cheetham 《Applied microbiology and biotechnology》1993,39(2):174-180
Transformed root tissue of Beta vulgaris (Detroit Dark Red) was permeabilized to stimulate the release of intracellularly stored betanin without adverse affects on tissue viability as measured by biomass accumulation. Product release of up to 15% (w/w) was achieved by heat treatment at 42°C for 45 min with minimal effect on viability. Higher levels of product release were obtained with increasing temperature and exposure, but at the expense of viability. Viability was measured by comparing dry weight increases of permeabilized tissue 3 days after treatment vs non-permeabilized tissue over the same time interval. Recovery of heat-treated tissue was improved by addition of CaCl2 (20 mm for 10 min) post-heat treatment. Betanin release up to 15% was also obtained at ambient temperature (25°C) by addition of up to 20 mm (NH4)2SO4 in the presence of 1 mm ethylenediaminetetraacetic acid (EDTA).
Correspondence to: A. A. DiIorio 相似文献
90.
Jrn Elsner Johannes Norgauer Gustav J. Dobos Andreas Emmendrffer Erwin Schpf Alexander Kapp Joachim Roesler 《Journal of cellular physiology》1993,157(3):637-643
Flow cytometric analyses were performed to study intracellular single-cell calcium transients ([Ca2+]i) in suspended human neutrophils during the initial phase of N-formyl peptide stimulation. Thereby, two neutrophil populations became apparent. Early maximally Ca2+-responding (high fluorescence) neutrophils and not-yet Ca2+-responding (low fluorescence) neutrophils, but no neutrophils with intermediate levels of [Ca2+]i, were detected. Within 7 s the number of low fluorescence neutrophils decreased and the number of high fluorescence neutrophils increased maximally. This suggests that [Ca2+]i transients occurred abruptly in individual neutrophils within a time interval below 1 s. At lower N-formyl peptide concentrations the lag times of individual neutrophils and the interval time of maximal activation of the [Ca2+]i-responding neutrophil population increased, however the percentage of [Ca2+]i-responding cells decreased. Surprisingly, no influence of the N-formyl peptide concentration on the [Ca2+]i-induced fluorescence signal of the individual cell was observed: it was always in an almost maximal range or not responding. In parallel, binding studies performed with fluorescein-labeled N-formyl peptide revealed that the heterogeneity of [Ca2+]i-responding cells cannot be explained by different receptor occupancy. In summary, this study demonstrates that [Ca2+]i transients induced by N-formyl peptides in suspended individual human neutrophils occur very rapidly in an almost “all-or-none manner” and that the mean increasing fluorescence signal of a calcium indicator within a whole neutrophil population results from varying lag times of the individual cells, rather than from the mean simultaneous progress of many cells. © 1993 Wiley-Liss, Inc. 相似文献