全文获取类型
收费全文 | 1596篇 |
免费 | 195篇 |
国内免费 | 1篇 |
出版年
2022年 | 13篇 |
2021年 | 26篇 |
2020年 | 25篇 |
2019年 | 23篇 |
2018年 | 26篇 |
2017年 | 29篇 |
2016年 | 34篇 |
2015年 | 61篇 |
2014年 | 60篇 |
2013年 | 69篇 |
2012年 | 92篇 |
2011年 | 97篇 |
2010年 | 62篇 |
2009年 | 56篇 |
2008年 | 79篇 |
2007年 | 74篇 |
2006年 | 67篇 |
2005年 | 63篇 |
2004年 | 69篇 |
2003年 | 55篇 |
2002年 | 50篇 |
2001年 | 51篇 |
2000年 | 33篇 |
1999年 | 37篇 |
1998年 | 17篇 |
1997年 | 17篇 |
1996年 | 18篇 |
1995年 | 20篇 |
1994年 | 16篇 |
1993年 | 13篇 |
1992年 | 45篇 |
1991年 | 35篇 |
1990年 | 30篇 |
1989年 | 26篇 |
1988年 | 22篇 |
1987年 | 15篇 |
1986年 | 23篇 |
1985年 | 20篇 |
1984年 | 20篇 |
1983年 | 16篇 |
1982年 | 17篇 |
1979年 | 17篇 |
1978年 | 11篇 |
1977年 | 11篇 |
1975年 | 8篇 |
1974年 | 13篇 |
1973年 | 10篇 |
1972年 | 8篇 |
1971年 | 11篇 |
1970年 | 9篇 |
排序方式: 共有1792条查询结果,搜索用时 15 毫秒
91.
Priyamvada Acharya Timothy S. Luongo Mark K. Louder Krisha McKee Yongping Yang Young Do Kwon John R. Mascola Pascal Kessler Loïc Martin Peter D. Kwong 《Structure (London, England : 1993)》2013,21(6):1018-1029
- Download : Download high-res image (334KB)
- Download : Download full-size image
92.
Daisuke Tsuru Heizo Kira Takehiko Yamamoto Juichiro Fukumoto 《Bioscience, biotechnology, and biochemistry》2013,77(9):856-862
The neutral protease of Bacillus amylosacchariticus was inactivated by low concentrations of several metal-chelating agents and the inactivated enzyme with EDTA restored its activity almost completely by the addition of Zn++ or Co++ and partially by Fe++ or Mn++, if these metal ions were added shortly after the EDTA-treatment. The native enzyme was found to contain 0.19% of zinc together with a significant amount of calcium. Parallel increase in specific activity and zinc content of enzyme preparation was observed throughout the purification procedure. The elution pattern of enzyme activity on a CM-cellulose column chromatography also completely coincided with that of protein-bound zinc. A zinc-free inactive enzyme was also reactivated by the addition of zinc or cobalt ions, clearly showing that the neutral protease of B. amylosacchariticus is a zinc mctalloenzyme. 相似文献
93.
Daisuke Tsuru Heizo Kira Takehiko Yamamoto Juichiro Fukumoto 《Bioscience, biotechnology, and biochemistry》2013,77(3):330-335
Some physicochemical properties and amino acid composition of the alkaline protease of B. amylosacchariticus were determined. The molecular weight and sedimentation coefficient were estimated to be 22,700 and 2.89 s, respectively, and the amino terminal amino acid was identified to be alanine. The enzyme contained 15.9% of nitrogen and was composed of 220 residues of amino acid: lys6, his5, arg3, asp20, thr14, ser37, glu12, pro10, gly25 ala27 val20, met3, isoleu12, leu12, tyr9, phe2, try3 and amide ammonia16 The results indicate that protein nature and chemical properties of the alkaline protease presented here are distinct from those of alkaline proteases obtained from the other strains of B. subtilis, such as subtilopeptidase A, B and BPN’ 相似文献
94.
Rafael Luis Kessler Daniela Fiori Gradia Rita de Cássia Pontello Rampazzo édio Elígio Louren?o Nilson José Fidêncio Lauro Manhaes Christian Macagnan Probst Andréa Rodrigues ávila Stenio Perdig?o Fragoso 《PloS one》2013,8(6)
Trypanosoma cruzi is the etiological agent of Chagas disease, an illness that affects about 10 million people, mostly in South America, for which there is no effective treatment or vaccine. In this context, transgenic parasites expressing reporter genes are interesting tools for investigating parasite biology and host-parasite interactions, with a view to developing new strategies for disease prevention and treatment. We describe here the construction of a stably transfected fluorescent T. cruzi clone in which the GFP gene is integrated into the chromosome carrying the ribosomal cistron in T. cruzi Dm28c. This fluorescent T. cruzi produces detectable amounts of GFP only at replicative stages (epimastigote and amastigote), consistent with the larger amounts of GFP mRNA detected in these forms than in the non replicative trypomastigote stages. The fluorescence signal was also strongly correlated with the total number of parasites in T. cruzi cultures, providing a simple and rapid means of determining the growth inhibitory dose of anti-T.cruzi drugs in epimastigotes, by fluorometric microplate screening, and in amastigotes, by the flow cytometric quantification of T. cruzi-infected Vero cells. This fluorescent T. cruzi clone is, thus, an interesting tool for unbiased detection of the proliferating stages of the parasite, with multiple applications in the genetic analysis of T. cruzi, including analyses of host-parasite interactions, gene expression regulation and drug development. 相似文献
95.
Vera Y. Chung Rebecca Konietzny Philip Charles Benedikt Kessler Roman Fischer Benjamin W. Turney 《Fly》2016,10(2):91-100
Kidney stone disease is a major health burden with a complex and poorly understood pathophysiology. Drosophila Malpighian tubules have been shown to resemble human renal tubules in their physiological function. Herein, we have used Drosophila as a model to study the proteomic response to crystal formation induced by dietary manipulation in Malpighian tubules. Wild-type male flies were reared in parallel groups on standard medium supplemented with lithogenic agents: control, Sodium Oxalate (NaOx) and Ethylene Glycol (EG). Malpighian tubules were dissected after 2 weeks to visualize crystals with polarized light microscopy. The parallel group was dissected for protein extraction. A new method of Gel Assisted Sample Preparation (GASP) was used for protein extraction. Differentially abundant proteins (p<0.05) were identified by label-free quantitative proteomic analysis in flies fed with NaOx and EG diet compared with control. Their molecular functions were further screened for transmembrane ion transporter, calcium or zinc ion binder. Among these, 11 candidate proteins were shortlisted in NaOx diet and 16 proteins in EG diet. We concluded that GASP is a proteomic sample preparation method that can be applied to individual Drosophila Malpighian tubules. Our results may further increase the understanding of the pathophysiology of human kidney stone disease. 相似文献
96.
Midpoint attractors and species richness: Modelling the interaction between environmental drivers and geometric constraints
下载免费PDF全文
![点击此处可从《Ecology letters》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Robert K. Colwell Nicholas J. Gotelli Louise A. Ashton Jan Beck Gunnar Brehm Tom M. Fayle Konrad Fiedler Matthew L. Forister Michael Kessler Roger L. Kitching Petr Klimes Jürgen Kluge John T. Longino Sarah C. Maunsell Christy M. McCain Jimmy Moses Sarah Noben Katerina Sam Legi Sam Arthur M. Shapiro Xiangping Wang Vojtech Novotny 《Ecology letters》2016,19(9):1009-1022
We introduce a novel framework for conceptualising, quantifying and unifying discordant patterns of species richness along geographical gradients. While not itself explicitly mechanistic, this approach offers a path towards understanding mechanisms. In this study, we focused on the diverse patterns of species richness on mountainsides. We conjectured that elevational range midpoints of species may be drawn towards a single midpoint attractor – a unimodal gradient of environmental favourability. The midpoint attractor interacts with geometric constraints imposed by sea level and the mountaintop to produce taxon‐specific patterns of species richness. We developed a Bayesian simulation model to estimate the location and strength of the midpoint attractor from species occurrence data sampled along mountainsides. We also constructed midpoint predictor models to test whether environmental variables could directly account for the observed patterns of species range midpoints. We challenged these models with 16 elevational data sets, comprising 4500 species of insects, vertebrates and plants. The midpoint predictor models generally failed to predict the pattern of species midpoints. In contrast, the midpoint attractor model closely reproduced empirical spatial patterns of species richness and range midpoints. Gradients of environmental favourability, subject to geometric constraints, may parsimoniously account for elevational and other patterns of species richness. 相似文献
97.
Monensin is a natural antibiotic that exhibits high affinity to certain metal ions. In order to explore its potential in coordination chemistry, circular dichroism (CD) spectra of monensic acid A (MonH) and its derivatives containing monovalent cations (Li+, Na+, K+, Rb+, Ag+, and Et4N+) in methanolic solutions were measured and compared to computational models. Whereas the conventional CD spectroscopy allowed recording of the transitions down to 192 nm, synchrotron radiation circular dichroism (SRCD) revealed other bands in the 178–192 nm wavelength range. CD signs and intensities significantly varied in the studied compounds, in spite of their similar crystal structure. Computational modeling based on the Density Functional Theory (DFT) and continuum solvent model suggests that the solid state monensin structure is largely conserved in the solutions as well. Time‐dependent Density Functional Theory (TDDFT) simulations did not allow band‐to‐band comparison with experimental spectra due to their limited precision, but indicated that the spectral changes were caused by a combination of minor conformational changes upon the monovalent cation binding and a direct involvement of the metal electrons in monensin electronic transitions. Both the experiment and simulations thus show that the CD spectra of monensin complexes are very sensitive to the captured ions and can be used for their discrimination. Chirality 28:420–428, 2016. © 2016 Wiley Periodicals, Inc. 相似文献
98.
99.
Mart Krupovic Kira S. Makarova Yuri I. Wolf Sofia Medvedeva David Prangishvili Patrick Forterre Eugene V. Koonin 《Environmental microbiology》2019,21(6):2056-2078
To explore the diversity of mobile genetic elements (MGE) associated with archaea of the phylum Thaumarchaeota, we exploited the property of most MGE to integrate into the genomes of their hosts. Integrated MGE (iMGE) were identified in 20 thaumarchaeal genomes amounting to 2 Mbp of mobile thaumarchaeal DNA. These iMGE group into five major classes: (i) proviruses, (ii) casposons, (iii) insertion sequence-like transposons, (iv) integrative-conjugative elements and (v) cryptic integrated elements. The majority of the iMGE belong to the latter category and might represent novel families of viruses or plasmids. The identified proviruses are related to tailed viruses of the order Caudovirales and to tailless icosahedral viruses with the double jelly-roll capsid proteins. The thaumarchaeal iMGE are all connected within a gene sharing network, highlighting pervasive gene exchange between MGE occupying the same ecological niche. The thaumarchaeal mobilome carries multiple auxiliary metabolic genes, including multicopper oxidases and ammonia monooxygenase subunit C (AmoC), and stress response genes, such as those for universal stress response proteins (UspA). Thus, iMGE might make important contributions to the fitness and adaptation of their hosts. We identified several iMGE carrying type I-B CRISPR-Cas systems and spacers matching other thaumarchaeal iMGE, suggesting antagonistic interactions between coexisting MGE and symbiotic relationships with the ir archaeal hosts. 相似文献
100.
The glycosomes of trypanosomatids are essential organelles that are evolutionarily related to peroxisomes of other eukaryotes. The peroxisomal RING proteins-PEX2, PEX10 and PEX12-comprise a network of integral membrane proteins that function in the matrix protein import cycle. Here, we describe PEX10 and PEX12 in Trypanosoma brucei, Leishmania major, and Trypanosoma cruzi. We expressed GFP fusions of each T. brucei coding region in procyclic form T. brucei, where they localized to glycosomes and behaved as integral membrane proteins. Despite the weak transmembrane predictions for TbPEX12, protease protection assays demonstrated that both the N and C termini are cytosolic, similar to mammalian PEX12. GFP fusions of T. cruzi PEX10 and L. major PEX12 also localized to glycosomes in T. brucei indicating that glycosomal membrane protein targeting is conserved across trypanosomatids. 相似文献