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141.
The reaction of ribose with horseradish peroxidase in the presence of H2O2 is accompanied by light emission. The detection of horseradish peroxidase Compound II (FeO2+) indicates that the enzyme participates in a normal peroxidatic cycle. Hydrogen peroxide converts horseradish peroxidase into Compound I (FeO3+) which in turn is converted into Compound II by abstracting a hydrogen atom from ribose forming a ribosyl radical. In aerated solutions oxygen rapidly adds to the ribosyl radical. Based on the spectral characteristics and the enhancement of the chemiluminescence by chlorophyll-a, xanthene dyes, D2O and DABCO, it is suggested that the excited species, apparently triplet carbonyls and 1O2, are formed from the bimolecular decay of the peroxyl radicals via the Russell mechanism. 相似文献
142.
143.
Localization of the receptor gene for type D simian retroviruses on human chromosome 19. 总被引:15,自引:9,他引:6 下载免费PDF全文
M A Sommerfelt B P Williams A McKnight P N Goodfellow R A Weiss 《Journal of virology》1990,64(12):6214-6220
Simian retrovirus (SRV) serotypes 1 to 5 are exogenous type D viruses causing immune suppression in macaque monkeys. These viruses exhibit receptor interference with each other, with two endogenous type D viruses of the langur (PO-1-Lu) and squirrel monkey, and with two type C retroviruses, feline endogenous virus (RD114/CCC) and baboon endogenous virus (BaEV), indicating that each utilizes the same cell surface receptor (M. A. Sommerfelt and R. A. Weiss, Virology 176:58-69, 1990). Vesicular stomatitis virus pseudotype particles bearing envelope glycoproteins of RD114, BaEV, and the seven SRV strains were employed to detect receptors expressed in human-rodent somatic cell hybrids segregating human chromosomes. The only human chromosome common to all the susceptible hybrids was chromosome 19. By using hybrids retaining different fragments of chromosome 19, a provisional subchromosomal localization of the receptor gene was made to 19q13.1-13.2. Antibodies previously reported to be specific to a BaEV receptor (L. Thiry, J. Cogniaux-Leclerc, R. Olislager, S. Sprecher-Goldberger, and P. Burkens, J. Virol. 48:697-708, 1983) did not block BaEV, RD114, or SRV pseudotypes or syncytia. Antibodies to known surface markers determined by genes mapped to chromosome 19 did not block virus-receptor interaction. The identity of the receptor remains to be determined. 相似文献
144.
Biological and molecular variability of human immunodeficiency virus type 2 isolates from The Gambia. 总被引:14,自引:9,他引:5 下载免费PDF全文
T F Schulz D Whitby J G Hoad T Corrah H Whittle R A Weiss 《Journal of virology》1990,64(10):5177-5182
Seven new human immunodeficiency virus type 2 (HIV-2) isolates (CBL-20 to CBL-26) from The Gambia were characterized. Their cytopathogenicity and growth in vitro correlated with the severity of clinical disease. CBL-22 was highly sensitive to neutralization by HIV-2 sera and was cross-neutralized by some HIV-1 sera. These findings, the differing sizes of envelope glycoproteins of individual isolates, and the sequence analysis of amplified regions of the viral DNAs show that these HIV-2 isolates from one geographical region in West Africa exhibit biological and genome variability comparable to that observed for HIV-1. 相似文献
145.
Clinical and biochemical studies in three patients with severe early infantile Cockayne syndrome 总被引:2,自引:0,他引:2
Jaak Jacken Helmut Klocker Helga Schwaiger Romuald Bellmann Monica Hirsch-Kauffmann Manfred Schweiger 《Human genetics》1989,83(4):339-346
Summary We present clinical and biochemical data from three patients with severe Cockayne syndrome (CS) of very early onset. Unlike in classic CS, signs became evident in the first weeks of life and led to unusually early death. Fibroblasts from two of the patients showed a complete defect of the repair of UV-induced thymine dimer lesions. They were unable to remove thymine dimer lesions from their DNA, had a severe reduction of the RNA synthesis rates after UV irradiation, and showed no reactivation of an UV-inactivated indicator gene and no DNA recondensation after UV irradiation. DNA repair investigated in these two fibroblast cell strains resembled that of xeroderma pigmentosum cells of complementation group A. In contrast, fibroblasts from the third patient showed the same in vitro repair characteristics as classic CS cells. 相似文献
146.
Helmut Wieczorek Gabriele Wolff 《Journal of comparative physiology. A, Neuroethology, sensory, neural, and behavioral physiology》1989,164(6):825-834
1. | The electrophysiology of the sugar receptor in labellar taste hairs ofDrosophila melanogaster (Diptera) was investigated using 33 monosaccharides, oligosaccharides, glucosides, and amino acids which in calyptrate flies are known to bind to specific receptor sites or which may be assigned to specific receptor sites on the basis of structural criteria. |
2. | The pyranose site ofDrosophila is very similar to the pyranose site of the calyptrate flies: regarding monosaccharides of the pyranose type three adjacent equatorial hydroxyl groups (C-2, C-3, C-4) seem to be important for stimulating effectiveness. On the other hand, it exhibits a more rigid stereospecificity with regard to the substituents at C-1 and C-5. |
3. | A furanose site as in calyptrate flies does not exist inDrosophila. First, D-galactose, phenylalanine and 2,5-anhydro-D-mannitol are not or nearly not stimulatory. Secondly, according to different sugar receptor responses after treatment of the taste hairs with papain, D-fucose binds to another receptor site than D-fructose. Thirdly, the effective conformation of D-fructose is not the furanose, but most probably the pyranose form as can be concluded from experiments with freshly prepared and equilibrium solutions of D-fructose. |
4. | The characteristic differences between the properties of the sugar receptors ofDrosophila and of the calyptrate flies lead to the suggestion that the actual number of types of receptor sites in the various fly species is greater than assumed up till now. The broad specificity of the sugar receptors of flies may therefore result from a mosaic of different types of highly specific receptor sites. |
147.
A prokaryotic vector, pGE374, containing the recA and lacZ genes, out-of-frame, was used for the expression of cDNA derived from the putative polymerase-encoding gene of the coronavirus mouse hepatitis virus strain A59 (MHV-A59). The pGE374/viral recombinant vector generates a tripartite bacterial/viral protein composed of a segment of the RecA protein at the N terminus, the coronaviral sequences in the middle, and an enzymatically active beta-galactosidase at the C terminus. Rabbits immunized with such recombinant proteins generated antibodies to the MHV-A59 portion of the tripartite protein. Because the MHV-A59 polymerase proteins have been difficult to identify during infection, we used a novel method to demonstrate the viral specificity of the antiserum. The viral cDNA was excised from the expression vector, and transferred to a pGem vector, downstream from and in-frame with a portion of the cat gene. This construct contained a bacteriophage RNA polymerase promoter that enabled the cell-free synthesis of a fusion protein that was used to verify that antibodies were generated to the expressed viral DNA. This strategy was shown to successfully result in the specific generation of antibodies to the encoded information of the viral cDNA. Furthermore, this method has general applicability in the generation and characterization of antibodies directed against proteins encoded in cDNAs. 相似文献
148.
Ammonia assimilation pathways in nitrogen-fixing Clostridium kluyverii and Clostridium butyricum. 总被引:2,自引:1,他引:1 下载免费PDF全文
Pathways of ammonia assimilation into glutamic acid were investigated in ammonia-grown and N2-fixing Clostridium kluyverii and Clostridium butyricum by measuring the specific activities of glutamate dehydrogenase, glutamine synthetase, and glutamate synthase. C. kluyverii had NADPH-glutamate dehydrogenase with a Km of 12.0 mM for NH4+. The glutamate dehydrogenase pathway played an important role in ammonia assimilation in ammonia-grown cells but was found to play a minor role relative to that of the glutamine synthetase/NADPH-glutamate synthase pathway in nitrogen-fixing cells when the intracellular NH4+ concentration and the low affinity of the enzyme for NH4+ were taken into account. In C. butyricum grown on glucose-salt medium with ammonia or N2 as the nitrogen source, glutamate dehydrogenase activity was undetectable, and the glutamine synthetase/NADH-glutamate synthase pathway was the predominant pathway of ammonia assimilation. Under these growth conditions, C. butyricum also lacked the activity of glucose-6-phosphate dehydrogenase, which catalyzes the regeneration of NADPH from NADP+. However, high activities of glucose-6-phosphate dehydrogenase as well as of NADPH-glutamate dehydrogenase with a Km of 2.8 mM for NH4+ were present in C. butyricum after growth on complex nitrogen and carbon sources. The ammonia-assimilating pathway of N2-fixing C. butyricum, which differs from that of the previously studied Bacillus polymyxa and Bacillus macerans, is discussed in relation to possible effects of the availability of ATP and of NADPH on ammonia-assimilating pathways. 相似文献
149.
Function of a heterologous muscarinic receptor in T cell antigen receptor signal transduction mutants 总被引:7,自引:0,他引:7
M A Goldsmith D M Desai T Schultz A Weiss 《The Journal of biological chemistry》1989,264(29):17190-17197
Previously we have described a system of somatic cell genetics (J.CaM1 and J.CaM2) for analyzing signal transduction via the T cell antigen receptor complex (CD3/Ti). Here we describe a third mutant, J.CaM3, which also expresses high levels of receptors that are functionally impaired. Like J.CaM1, J.CaM3 demonstrates partial signal transduction via CD3/Ti to only certain stimuli. J.CaM1, J.CaM2, and J.CaM3 define three non-Ti complementation groups involved in receptor function. To evaluate the mutations further we have introduced a heterologous receptor, the human muscarinic receptor 1 (HM1), into the parental Jurkat and mutant cell lines. This receptor demonstrates signal transduction competence in all these hosts, indicating that 1) T cells express the necessary apparatus for the coupling of HM1 to second messenger generation and 2) the mutations in the J.CaM family all affect molecules that are specific to CD3/Ti, and not HM1, function. Finally, the HM1 receptor exhibits partial sensitivity to cholera toxin in Jurkat cells, in contrast to the virtually complete sensitivity of CD3/Ti to cholera toxin. 相似文献
150.
An antibody against secretogranin I (chromogranin B) is packaged into secretory granules 总被引:16,自引:8,他引:8 下载免费PDF全文
P Rosa U Weiss R Pepperkok W Ansorge C Niehrs E H Stelzer W B Huttner 《The Journal of cell biology》1989,109(1):17-34
We have investigated the sorting and packaging of secretory proteins into secretory granules by an immunological approach. An mAb against secretogranin I (chromogranin B), a secretory protein costored with various peptide hormones and neuropeptides in secretory granules of many endocrine cells and neurons, was expressed by microinjection of its mRNA into the secretogranin I-producing cell line PC12. An mAb against the G protein of vesicular stomatitis virus--i.e., against an antigen not present in PC12 cells--was expressed as a control. The intracellular localization and the secretion of the antibodies was studied by double-labeling immunofluorescence using the conventional and the confocal microscope, as well as by pulse-chase experiments. The secretogranin I antibody, like the control antibody, was transported along the secretory pathway to the Golgi complex. However, in contrast to the control antibody, which was secreted via the constitutive pathway, the secretogranin I antibody formed an immunocomplex with secretogranin I, was packaged into secretory granules, and was released by regulated exocytosis. Our results show that a constitutive secretory protein, unaltered by genetic engineering, can be diverted to the regulated pathway of secretion by its protein-protein interaction with a regulated secretory protein. The data also provide the basis for immunologically studying the role of luminally exposed protein domains in the biogenesis and function of regulated secretory vesicles. 相似文献