全文获取类型
收费全文 | 100631篇 |
免费 | 782篇 |
国内免费 | 887篇 |
专业分类
102300篇 |
出版年
2023年 | 80篇 |
2022年 | 114篇 |
2021年 | 195篇 |
2020年 | 154篇 |
2019年 | 175篇 |
2018年 | 11987篇 |
2017年 | 10789篇 |
2016年 | 7744篇 |
2015年 | 998篇 |
2014年 | 742篇 |
2013年 | 834篇 |
2012年 | 4902篇 |
2011年 | 13345篇 |
2010年 | 12372篇 |
2009年 | 8544篇 |
2008年 | 10216篇 |
2007年 | 11790篇 |
2006年 | 667篇 |
2005年 | 849篇 |
2004年 | 1286篇 |
2003年 | 1308篇 |
2002年 | 1020篇 |
2001年 | 325篇 |
2000年 | 213篇 |
1999年 | 81篇 |
1998年 | 82篇 |
1997年 | 72篇 |
1996年 | 64篇 |
1995年 | 60篇 |
1994年 | 55篇 |
1993年 | 67篇 |
1992年 | 56篇 |
1991年 | 67篇 |
1990年 | 24篇 |
1989年 | 30篇 |
1988年 | 45篇 |
1987年 | 28篇 |
1986年 | 19篇 |
1985年 | 21篇 |
1984年 | 34篇 |
1983年 | 34篇 |
1982年 | 18篇 |
1981年 | 29篇 |
1980年 | 21篇 |
1979年 | 20篇 |
1974年 | 17篇 |
1973年 | 17篇 |
1972年 | 251篇 |
1971年 | 281篇 |
1962年 | 24篇 |
排序方式: 共有10000条查询结果,搜索用时 0 毫秒
81.
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
82.
H-2 loss variant sublines of a sarcoma (M-AS), induced by methylcholanthrene in an (A × A.SW)F1 mouse, were used to study the role of the MHC products in the recognition of MC-TSTA. The two reciprocal variant sublines (M-A and M-S) were found to express the TSTA of the original tumor as shown by cross-reactions in graft rejection experiments performed in (A × A.SW)F1 mice. In the A/Sn and A.SW mice the presence of the reciprocal parental H-2 antigens on the immunizing cells decreased the response against the tumor antigens. An admixture of lymphocytes derived from hyperimmune mice inhibited the outgrowth of the tumor cells. The growth inhibition was mediated by T cells and was H-2 restricted. Cells derived from hyperimmune syngeneic mice inhibited the outgrowth of the variant subline used for immunization but had no effect on the reciprocal variant subline. 相似文献
83.
This paper reviews the systematic status and geographical distribution of 30 odd species of freshwater calanoid copepods in Southeast Asia. Three regions are delineated based on the diversity and abundance of these organisms; Region I being the Southasiatic element, Region II the attenuated belt of copepods and Region III rich in endemic species. The implication of a north-south diminishing of copepod species to inland fisheries is postulated. 相似文献
84.
The contribution of the 1–6 N-terminal sequence to the conformational properties of the S-peptide (the 1–20 sequence of ribonuclease A) was assessed by determining in the ribonuclease S′ system the helical content and the binding capability of synthetic [Orn10]-S-peptide analogs, in which lysine1, glutamic2 and threonine3 were progressively deleted, alanine4 and alanine5were alternatively replaced by serine, and alanine6 was substituted by serine or proline. Both the deletion of the three N-terminal residues and the alanine6/proline replacement produces the loss of the helical structure up to lysine7. No or minor effects are found in all other cases. From the comparison of the binding data, the energy for the conformational stabilization of the N-terminal region was calculated to amount to 1.4 kcal/mol. The results are discussed in comparison with the known x-ray data of the enzyme, with some predictive rules of secondary structure which were applied to this region and with the known phylogenetic variance of the residues in this region. 相似文献
85.
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
86.
Pablo Fernando Caligiore-Gei Pedro Della-Gaspera Eliana Benitez Christian Tarnowski 《The Plant Pathology Journal》2022,38(4):296
The cucurbit powdery mildew (CPM) caused by different fungal species is a major concern for cucurbit crops around the world. In Argentina CPM constitutes the most common and damaging disease for cucurbits, especially for squash crops (Cucurbita moschata). The present study displays initial insights into the knowledge of the disease in western Argentina, including the determination of the prevalent species causing CPM, as well as the evaluation of the resistance of squash cultivars and breeding lines. Fungal colonies were isolated from samples collected in Mendoza province, Argentina. A field trial was also performed to assess the resistance of five squash accessions, including commercial cultivars and breeding lines. The severity of CPM was analyzed and epidemiological models were built based on empirical data. The morphological determinations and analysis with specific molecular markers confirmed Podosphaera xanthi as the prevalent causal agent of CPM in Mendoza. The results od the field trial showed differences in the resistance trait among the squash accessions. The advanced breeding line BL717/1 showed promising results as source of CPM resistance for the future development of open pollinated resistant cultivars, a crucial tool for an integrative control of the disease. 相似文献
87.
Staining of Some Specific Regions of Human Chromosomes,particularly the Secondary Constriction of No. 9 总被引:23,自引:0,他引:23
SEVERAL procedures have been described recently which produce specific patterns of differential staining in human chromosomes1–9. Techniques which involve DNA denaturation and reannealing reveal deeply stained areas on centromere and secondary constriction regions which have been equated with constitutive heterochromatin9. 相似文献
88.
The biochemical mechanisms underlying thidiazuron (TDZ)-induced regeneration in plant cells have not been clearly elucidated.
Exposure of leaf explants of Echinacea purpurea to a medium containing TDZ results in undifferentiated cell proliferation and differentiated growth as mixed shoot organogenesis
and somatic embryogenesis. The current studies were undertaken to determine the potential roles of auxin, indoleamines, and
ion signaling in the dedifferentiation and redifferentiation of plant cells. E. purpurea leaf explants were found to contain auxin and the related indoleamine neurotransmitters, melatonin, and serotonin. The levels
of these endogenous indoleamines were increased by exposure to TDZ associated with the induction of regeneration. The auxin-transport
inhibitor 2,3,5-triiodobenzoic acid and auxin action inhibitor, p-chlorophenoxyisobutyric acid decreased the TDZ-induced regeneration but increased concentrations of endogenous serotonin
and melatonin. As well, inhibitors of calcium and sodium transport significantly reduced TDZ-induced morphogenesis while increasing
endogenous indoleamine content. These data indicate that TDZ-induced regeneration is the manifestation of a metabolic cascade
that includes an initial signaling event, accumulation, and transport of endogenous plant signals such as auxin and melatonin,
a system of secondary messengers, and a concurrent stress response. 相似文献
89.
90.
Carlos?Talavera Fernando?Contreras Francisco?Espadas Gabriela?Fuentes J.?M.?SantamaríaEmail author 《Plant Cell, Tissue and Organ Culture》2005,83(3):287-292
Plantlets of coconut were cultured in vitro under three different ambient conditions including a standard culture room, a culture room inside a glasshouse with natural light but controlled temperature, and a standard glasshouse with natural light and natural fluctuations of temperature. Plantlets from the 3 treatments were compared in terms of growth, plant survival as well as net photosynthesis and efficiency of PSII (Fv/Fm ratio) both at the end of the in vitro stage and at 3 stages of ex vitro acclimatization. At the end of the in vitro stage, plantlets cultured in vitro under glasshouse conditions showed the best performance showing the highest photosynthesis rate, dry weight and number of leaves. Plantlets from the standard culture room showed the lowest photosynthesis and growth rate. After 6 months of ex vitro acclimatization, plantlets originally grown in vitro under glasshouse conditions maintained better field survival and growth rates in terms of fresh weight, dry weight and leaf number than plantlets originally grown in vitro in the standard culture room. Although more studies are required to define the reason for this effect, it is clear that the conditions of standard culture rooms are not the best for in vitro cultivation of coconut and perhaps other tropical species. 相似文献