全文获取类型
收费全文 | 103211篇 |
免费 | 1193篇 |
国内免费 | 1795篇 |
专业分类
106199篇 |
出版年
2024年 | 34篇 |
2023年 | 147篇 |
2022年 | 287篇 |
2021年 | 486篇 |
2020年 | 360篇 |
2019年 | 441篇 |
2018年 | 12223篇 |
2017年 | 10920篇 |
2016年 | 7871篇 |
2015年 | 1270篇 |
2014年 | 1099篇 |
2013年 | 1184篇 |
2012年 | 5180篇 |
2011年 | 13616篇 |
2010年 | 12507篇 |
2009年 | 8593篇 |
2008年 | 10282篇 |
2007年 | 11766篇 |
2006年 | 640篇 |
2005年 | 872篇 |
2004年 | 1303篇 |
2003年 | 1372篇 |
2002年 | 1053篇 |
2001年 | 434篇 |
2000年 | 306篇 |
1999年 | 195篇 |
1998年 | 102篇 |
1997年 | 89篇 |
1996年 | 64篇 |
1995年 | 68篇 |
1994年 | 67篇 |
1993年 | 71篇 |
1992年 | 79篇 |
1991年 | 88篇 |
1990年 | 41篇 |
1989年 | 39篇 |
1988年 | 53篇 |
1987年 | 32篇 |
1986年 | 27篇 |
1985年 | 23篇 |
1984年 | 33篇 |
1983年 | 32篇 |
1982年 | 16篇 |
1975年 | 16篇 |
1973年 | 21篇 |
1972年 | 258篇 |
1971年 | 282篇 |
1965年 | 20篇 |
1962年 | 31篇 |
1956年 | 15篇 |
排序方式: 共有10000条查询结果,搜索用时 0 毫秒
71.
Function of the <Emphasis Type="Italic">rel</Emphasis> Gene in <Emphasis Type="Italic">Escherichia coli</Emphasis> 总被引:6,自引:0,他引:6
Sokawa et al. suggest that rel- strains of Escherichia coli possess abnormal protein synthesizing machinery, which cannot carry out normal protein synthesis when the supply of amino-acids is limited. 相似文献
72.
AVRAM HERSHKO PIERRE MAMONT ROBERT SHIELDS GORDON M. TOMKINS 《Nature: New biology》1971,232(33):206-211
A hypothesis has been developed to relate stringent control in bacteria to a set of interactions involved in the regulation of growth of transformed and untransformed mammalian cells. 相似文献
73.
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
74.
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
75.
76.
【目的】葡聚糖酶是饲用添加剂的重要成分,本研究旨在从湖羊消化道微生物中挖掘性质优良的GH9家族葡聚糖酶基因,用于研发新型饲用酶制剂。【方法】从湖羊瘤胃微生物cDNA中扩增IDSGLUC9-25基因,在大肠杆菌中进行异源表达,对重组蛋白进行诱导表达和纯化,研究重组蛋白的酶学性质和底物水解模式。【结果】IDSGLUC9-25基因编码527个氨基酸,包含一个CelD_N结构和一个GH9家族催化结构域;重组蛋白rIDSGLUC9-25分子量约为62.7 kDa,最适反应温度和pH分别为40℃和6.0,在30-50℃下活性较高,在pH 4.0-8.0范围内能够保持较高的稳定性,经pH 4.0-8.0缓冲液处理1 h后残余活性均大于90%;底物谱分析表明,rIDSGLUC9-25能催化大麦β-葡聚糖、苔藓地衣多糖、魔芋胶和木葡聚糖,比活性分别为(443.55±24.48)、(65.56±5.98)、(122.37±2.85)和(159.16±7.73) U/mg;利用薄层色谱法(thin layer chromatography, TLC)和高效液相色谱法(high performance liquid chromatography, HPLC)分析水解产物发现,rIDSGLUC9-25降解大麦葡聚糖主要生成纤维三糖(占总还原糖64.19%±1.19%)和纤维四糖(占总还原糖26.24%±0.12%),催化地衣多糖主要生成纤维三糖(占总还原糖78.46%±0.89%)。【结论】本研究报道了一种来自密螺旋体属细菌的内切β-1,4-葡聚糖酶IDSGLUC9-25 (EC 3.2.1.4),能高效催化多糖底物生成纤维三糖和纤维四糖,为研发饲用酶制剂和制备低聚寡糖建立基础。 相似文献
77.
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
78.
79.
Barley seeds were treated for 3 h at 25°C with 240 mM ethyl methanesulphonate (EMS), washed for 18 h, treated with various concentrations of unbuffered sodium azide (pH 6.7–7.3) for 3 h at 25°C, re-dried to 30% water content and either sown immediately or stored at 25°C for 12 days and then sown. The synergistic action of sodium azide post-treatment has been demonstrated only for the EMS-induced M1 germination reduction, while the EMS-induced M1 sterility and the yield of M2 chlorophyll mutants were unaffected. The ?storage” recovery from EMS-induced mutagenic effects was insensitive to sodium azide post-treatment. The 12 day-seed storage at 25°C brought about an improvement of M1 germination, M1 survival, M1 fertility and a decrease in the amount of M2 mutants, regardless of whether sodium azide post-treatment was applied or not. 相似文献
80.
Guno Haskå 《Microbial ecology》1975,1(1):234-245
Myxobacteria presumably produce extracellular bacteriolytic enzymes when they are growing in soil. In order to study their ecological significance, adsorption experiments were performed with lytic enzymes produced byMyxococcus virescens in casitone media. Different soils as well as montmorillonite and kaolinite can rapidly adsorb the bacteriolytic but not the proteolytic enzymes. About 1 gm of montmorillonite per liter of cell-free culture solution is enough for the adsorption of 97% of the bacteriolytic enzymes. The adsorption per unit weight is about 100 times greater on montmorillonite than on kaolinite. About 40% of the adsorbed enzymes can be eluted with solutions of high pH or high ionic strength. The only desorbed bacteriolytic enzyme is the alanyl-∈-N-lysine endopeptidase. 相似文献