全文获取类型
收费全文 | 99005篇 |
免费 | 706篇 |
国内免费 | 889篇 |
专业分类
100600篇 |
出版年
2024年 | 17篇 |
2023年 | 56篇 |
2022年 | 110篇 |
2021年 | 231篇 |
2020年 | 134篇 |
2019年 | 165篇 |
2018年 | 11971篇 |
2017年 | 10786篇 |
2016年 | 7634篇 |
2015年 | 958篇 |
2014年 | 670篇 |
2013年 | 735篇 |
2012年 | 4739篇 |
2011年 | 13230篇 |
2010年 | 12247篇 |
2009年 | 8461篇 |
2008年 | 10054篇 |
2007年 | 11640篇 |
2006年 | 534篇 |
2005年 | 758篇 |
2004年 | 1215篇 |
2003年 | 1214篇 |
2002年 | 974篇 |
2001年 | 315篇 |
2000年 | 194篇 |
1999年 | 73篇 |
1998年 | 56篇 |
1997年 | 49篇 |
1996年 | 40篇 |
1995年 | 29篇 |
1994年 | 28篇 |
1993年 | 57篇 |
1992年 | 41篇 |
1991年 | 63篇 |
1990年 | 26篇 |
1989年 | 25篇 |
1988年 | 44篇 |
1987年 | 29篇 |
1984年 | 18篇 |
1983年 | 33篇 |
1982年 | 17篇 |
1981年 | 15篇 |
1980年 | 15篇 |
1978年 | 17篇 |
1976年 | 14篇 |
1975年 | 15篇 |
1972年 | 252篇 |
1971年 | 279篇 |
1965年 | 17篇 |
1962年 | 28篇 |
排序方式: 共有10000条查询结果,搜索用时 15 毫秒
71.
Kimball and Wilson1 reported that the arabinose analogue of cytidine (ara-C) inhibited DNA polymerase in a crude extract prepared from Ehrlich ascites cells. Furth and Cohen2 observed cytosine arabinoside triphosphate (ara-CTP) inhibited DNA polymerase in extracts from either calf thymus or bovine lymphosarcoma tissue, although these investigators3 had already found no effect of ara-CTP on DNA polymerase from Escherichia coli. The inhibition in both of these cases could be substantially reversed by dCTP; but incorporation of the arabinose nucleotide (ara-CMP) into DNA could not be unequivocally demonstrated. Graham and Whitmore4 reported the incorporation of ara-C into DNA in vivo and the inhibition of a DNA polymerase from L cells by ara-CTP. They found that ara-CMP was initially incorporated into small DNA strands but subsequently appeared in long strands. Momparler5 has presented evidence that, in vitro, ara-C incorporation was limited to the 3′-hydroxyl end of DNA chains. Such incorporation might be expected to block further chain elongation but this expectation was not supported by the evidence presented by Graham and Whitmore. 相似文献
72.
ALINA TAYLOR 《Nature: New biology》1971,234(48):144-145
JACOB and Fuerst1,2 demonstrated the presence of a bacteriolytic enzyme (λ-endolysin) in the induced cultures of lysogenic Escherichia coli K12 (λ). The enzyme was later identified as the product of gene R; of phage λ3 which is involved in bacterial lysis at the end of a latent period. The enzyme is apt to form spheroplast-like structures in E. coli2 and one would therefore expect its substrate to be murein. 相似文献
73.
74.
Heungsoon Felix Lee Roger Vivian Johnson 《Flexible Services and Manufacturing Journal》1991,3(2):91-120
We present a rough-cut analysis tool that quickly determines a few potential cost-effective designs at the initial design stage of flexible assembly systems (FASs) prior to a detailed analysis such as simulation. It uses quantitative methods for selecting and configuring the components of an FAS suitable for medium to high volumes of several similar products. The system is organized as a series of assembly stations linked with an automated material-handling system moving parts in a unidirectional flow. Each station consists of a single machine or of identical parallel machines. The methods exploit the ability of flexible hardware to switch almost instantaneously from product to product. Our approach is particularly suitable where the product mix is expected to be stable, since we combine the hardware-configuration phase with the task-allocation phase. For the required volume of products, we use integer programming to select the number of stations and the number of machines at each station and to allocate tasks to stations. We use queueing network analysis, which takes into account the mean and variance of processing times among different products to determine the necessary capacity of the material-handling system. We iterate between the two analyses to find the combined solution with the lowest costs. Work-in-process costs are also included in the analysis. Computational results are presented. 相似文献
75.
Antonio Balas Felix Garcia-Sanchez Fernando Gomez-Reino Jose L. Vicario 《Immunogenetics》1994,39(6):452-452
Correspondence to: J. L. Vicario. 相似文献
76.
Phosphorylation and dephosphorylation of ribosomal proteins have been suggested to participate in the regulation of protein synthesis in eukaryotic organisms. The present research focuses on the purification and partial characterization of a protein kinase from maize ribosomes that specifically phosphorylates acidic ribosomal proteins. Ribosomes purified from maize axes were used as the enzyme source. Purification of ribosomes was performed by centrifugation through a 0.5 M sucrose, 0.8 M KCl cushion. A protein kinase activity present in this fraction was released by extraction with 1.5 M KCl and further purified by diethylaminoethyl cellulose column chromatography. A peak containing protein kinase activity was eluted around 400 m M KCl. Analysis of this fraction by sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed one band of 38 kDa molecular mass, which cross-reacted in a western blot with antibodies raised against proteins from the large ribosomal subunit. This enzyme specifically phosphorylates one of the acidic ribosomal proteins (P2 ). Its activity is inhibited by Ca2+ and Zn2+ and is activated by Mg2+ , polylysine and spermine. The relevance of this protein kinase in reinitiating the protein synthesis process during germination is discussed. 相似文献
77.
78.
Perin L. Donnini M. Diomede L. Romano M. Tacconi M. T. Luisetti M. Salmona M. 《Cytotechnology》1991,7(1):25-32
An expression vector for G-CSF, pASLB3-3, was constructed and introduced into Namalwa KJM-1 cells (Hosoi et al., 1988), and cells resistant to 100 nM of methotrexate (MTX) were obtained. Among them, the highest producer, clone SC57, was selected and the productivity of this clone was further characterized. The maximal production of G-CSF was at the most 1.8 g/ml/day using a 25 cm2 tissue culture flask, even though the cell number was above 7×105 cells/ml. The limiting factors at high density were analyzed as the deficiency of nutrients, such as glucose, cysteine and serine, and pH control. The depression of specific G-CSF productivity per cell under the batch culture conditions was overcome by using a perfusion culture system, BiofermenterTM (Sato, 1983) with modifications of nutrients supplementation by a dialysis membrane and/or dissolved oxygen (DO) supplementation by microsilicone fibers. ITPSGF medium was modified to elevate concentrations of amino acids and glucose by 2.0- and 2.5-times, respectively. Under the control of pH at 7.4 and DO at 3 ppm, the specific G-CSF productivity was not depressed even at high cell density (above 1×107 cells/ml), and the amount of G-CSF reached 41 g/ml. These results indicated the possibility of finding the optimum culture conditions for the production of recombinant proteins by Namalwa KJM-1 cells.Abbreviations ABTS
2,2-Azino-di-(3-ethylbenzothiazoline)-6-sulfonic acid
- BSA
Bovine Serum Albumin
- BSA-PBS
Phosphate-buffered Saline without Ca2+ and Mg2+ containing Bovine Serum Albumin
- dhfr
Dihydrofolate Reductase
- DO
Dissolved Oxygen
- G-CSF
Granulocyte Colony-stimulating Factor
- HEPES
4-(2-Hydroxyethyl)-1-piperazineethansulfonic Acid
- IFN
Interferon
- MTX
Methotrexate
- PBS(-)
Phosphate-buffered saline without Ca2+ and Mg2+
- Tween-PBS
Phosphate-buffered saline without Ca2+ and Mg2+ containing 0.05% of Tween 20 相似文献
79.
Solid-phase peptide synthesis of somatostatin using mild base cleavage of N alpha-9-fluorenylmethyloxycarbonylamino acids 总被引:3,自引:0,他引:3
C D Chang A M Felix M H Jimenez J Meienhofer 《International journal of peptide and protein research》1980,15(5):485-494
Experimental details for the "Fmoc solid phase peptide synthesis" of somatostatin are described. The 9-fluorenylmethyloxycarbonyl group was rapidly and quantitatively cleaved by 55% piperidine in dimethylformamide and monitored (u.v.) manually. For a kinetic study, a centrifugal reactor with a photometric control system and reference cell was used at each stage. The symmetrical anhydride coupling reaction was rapid and either acetic anhydride or fluorescamine termination was incorporated to minimize formation of deletion peptides. Anchor-bond cleavage was effected with trifluoroacetic acid which simultaneously removed all the acid labile tert.-butyl side chain protecting groups. N alpha-9-fluorenylmethyloxycarbonyl peptides may be obtained by omitting the piperidine deprotection step after the last cycle of synthesis. From several syntheses, analytically pure di-S-protected somatostatin 14-peptide was obtained in 55-60% overall yield. The S-protecting groups were removed and the product was purified by gel filtration to give homogeneous dihydrosomatostatin (91%) yield. Oxidation of dihydrosomatostatin with potassium ferricyanide and purification by countercurrent distribution provided analytically pure homogeneous somatostatin. 相似文献
80.