全文获取类型
收费全文 | 2449篇 |
免费 | 222篇 |
专业分类
2671篇 |
出版年
2023年 | 20篇 |
2022年 | 38篇 |
2021年 | 48篇 |
2020年 | 40篇 |
2019年 | 58篇 |
2018年 | 80篇 |
2017年 | 63篇 |
2016年 | 99篇 |
2015年 | 135篇 |
2014年 | 143篇 |
2013年 | 155篇 |
2012年 | 221篇 |
2011年 | 197篇 |
2010年 | 147篇 |
2009年 | 106篇 |
2008年 | 138篇 |
2007年 | 111篇 |
2006年 | 114篇 |
2005年 | 82篇 |
2004年 | 95篇 |
2003年 | 93篇 |
2002年 | 81篇 |
2001年 | 37篇 |
2000年 | 35篇 |
1999年 | 27篇 |
1998年 | 17篇 |
1997年 | 16篇 |
1996年 | 16篇 |
1995年 | 14篇 |
1994年 | 17篇 |
1993年 | 16篇 |
1992年 | 13篇 |
1991年 | 15篇 |
1990年 | 13篇 |
1989年 | 19篇 |
1988年 | 18篇 |
1987年 | 12篇 |
1986年 | 9篇 |
1984年 | 7篇 |
1981年 | 10篇 |
1980年 | 6篇 |
1979年 | 8篇 |
1978年 | 6篇 |
1977年 | 9篇 |
1975年 | 8篇 |
1974年 | 10篇 |
1973年 | 8篇 |
1972年 | 5篇 |
1968年 | 5篇 |
1966年 | 6篇 |
排序方式: 共有2671条查询结果,搜索用时 15 毫秒
121.
John F. Darby Ewelina M. Krysztofinska Peter J. Simpson Aline C. Simon Pawel Leznicki Newran Sriskandarajah David S. Bishop Lisa R. Hale Caterina Alfano Maria R. Conte Santiago Martínez-Lumbreras Arjun Thapaliya Stephen High Rivka L. Isaacson 《PloS one》2014,9(11)
Background
The BAG6 complex resides in the cytosol and acts as a sorting point to target diverse hydrophobic protein substrates along their appropriate paths, including proteasomal degradation and ER membrane insertion. Composed of a trimeric complex of BAG6, TRC35 and UBL4A, the BAG6 complex is closely associated with SGTA, a co-chaperone from which it can obtain hydrophobic substrates.Methodology and Principal Findings
SGTA consists of an N-terminal dimerisation domain (SGTA_NT), a central tetratricopeptide repeat (TPR) domain, and a glutamine rich region towards the C-terminus. Here we solve a solution structure of the SGTA dimerisation domain and use biophysical techniques to investigate its interaction with two different UBL domains from the BAG6 complex. The SGTA_NT structure is a dimer with a tight hydrophobic interface connecting two sets of four alpha helices. Using a combination of NMR chemical shift perturbation, isothermal titration calorimetry (ITC) and microscale thermophoresis (MST) experiments we have biochemically characterised the interactions of SGTA with components of the BAG6 complex, the ubiquitin-like domain (UBL) containing proteins UBL4A and BAG6. We demonstrate that the UBL domains from UBL4A and BAG6 directly compete for binding to SGTA at the same site. Using a combination of structural and interaction data we have implemented the HADDOCK protein-protein interaction docking tool to generate models of the SGTA-UBL complexes.Significance
This atomic level information contributes to our understanding of the way in which hydrophobic proteins have their fate decided by the collaboration between SGTA and the BAG6 complex. 相似文献122.
BRL1 and BRL3 are novel brassinosteroid receptors that function in vascular differentiation in Arabidopsis 总被引:17,自引:0,他引:17
Caño-Delgado A Yin Y Yu C Vafeados D Mora-García S Cheng JC Nam KH Li J Chory J 《Development (Cambridge, England)》2004,131(21):5341-5351
Plant steroid hormones, brassinosteroids (BRs), are perceived by the plasma membrane-localized leucine-rich-repeat-receptor kinase BRI1. Based on sequence similarity, we have identified three members of the BRI1 family, named BRL1, BRL2 and BRL3. BRL1 and BRL3, but not BRL2, encode functional BR receptors that bind brassinolide, the most active BR, with high affinity. In agreement, only BRL1 and BRL3 can rescue bri1 mutants when expressed under the control of the BRI1 promoter. While BRI1 is ubiquitously expressed in growing cells, the expression of BRL1 and BRL3 is restricted to non-overlapping subsets of vascular cells. Loss-of-function of brl1 causes abnormal phloem:xylem differentiation ratios and enhances the vascular defects of a weak bri1 mutant. bri1 brl1 brl3 triple mutants enhance bri1 dwarfism and also exhibit abnormal vascular differentiation. Thus, Arabidopsis contains a small number of BR receptors that have specific functions in cell growth and vascular differentiation. 相似文献
123.
Metabolic engineering of isoprenoid biosynthesis in Arabidopsis for the production of taxadiene, the first committed precursor of Taxol 总被引:1,自引:0,他引:1
Besumbes O Sauret-Güeto S Phillips MA Imperial S Rodríguez-Concepción M Boronat A 《Biotechnology and bioengineering》2004,88(2):168-175
Paclitaxel (Taxol) is a widely used anticancer isoprenoid produced by the secondary metabolism of yew (Taxus sp.) trees. However, only limited amounts of Taxol or related metabolites (taxoids) can be obtained from the currently available sources. In this work we have taken the first step toward genetically engineering the biosynthesis of taxoids in angiosperms. The first committed step in Taxol biosynthesis is the production of taxadiene from geranylgeranyl diphosphate (GGPP), catalyzed by the plastid-localized enzyme taxadiene synthase (TXS). A recombinant T. baccata TXS lacking the putative plastid targeting peptide and fused to a C-terminal histidine (His) tag was shown to be enzymatically active in Escherichia coli. Constitutive production of the full-length His-tagged enzyme in Arabidopsis thaliana plants led to the accumulation of taxadiene and concomitant growth retardation and decreased levels of photosynthetic pigment in transgenic plants. Although these phenotypes may derive from a toxic effect of taxadiene, the lower accumulation of endogenous plastid isoprenoid products such as carotenoids and chlorophylls in transgenic plants also suggests that the constitutive production of an active TXS enzyme might alter the balance of the GGPP pool. Induction of transgene expression using a glucocorticoid-mediated system consistently resulted in a more efficient recruitment of GGPP for the production of taxadiene, which reached levels 30-fold higher than those in plants constitutively expressing the transgene. This accomplishment illustrates the possibility of engineering the production of taxoids and other GGPP-derived isoprenoids in crop plants despite the constraints associated with limited knowledge with regard to regulation of GGPP availability. 相似文献
124.
Sekhar A Santiago M Lam HN Lee JH Cavagnero S 《Protein science : a publication of the Protein Society》2012,21(7):1042-1055
Most known proteins have at least one local Hsp70 chaperone binding site. Does this mean that all proteins interact with Hsp70 as they fold? This study makes an initial step to address the above question by examining the interaction of the E.coli Hsp70 chaperone (known as DnaK) and its co-chaperones DnaJ and GrpE with a slow-folding E.coli substrate, RNase HD. Importantly, this protein is a nonobligatory client, and it is able to fold in vitro even in the absence of chaperones. We employ stopped-flow mixing, chromatography, and activity assays to analyze the kinetic perturbations induced by DnaK/DnaJ/GrpE (K/J/E) on the folding of RNase HD. We find that K/J/E slows down RNase HD''s apparent folding, consistent with the presence of transient chaperone-substrate interactions. However, kinetic retardation is moderate for this slow-folding client and it is expected to be even smaller for faster-folding substrates. Given that the interaction of folding-competent substrates such as RNase HD with the K/J/E chaperones is relatively short-lived, it does not significantly interfere with the timely production of folded biologically active substrate. The above mode of action is important because it preserves K/J/E bioavailability, enabling this chaperone system to act primarily by assisting the folding of other misfolded and (or) aggregation-prone cellular proteins that are unable to fold independently. When refolding is carried out in the presence of K/J and absence of the nucleotide exchange factor GrpE, some of the substrate population becomes trapped as a chaperone-bound partially unfolded state. 相似文献
125.
126.
Santiago?Comba Martín?Sabatini Simón?Menendez-Bravo Ana?ArabolazaEmail author Hugo?GramajoEmail author 《Biotechnology for biofuels》2014,7(1):172
Background
Microbial lipid production represents a potential alternative feedstock for the biofuel and oleochemical industries. Since Escherichia coli exhibits many genetic, technical, and biotechnological advantages over native oleaginous bacteria, we aimed to construct a metabolically engineered E. coli strain capable of accumulating high levels of triacylglycerol (TAG) and evaluate its neutral lipid productivity during high cell density fed-batch fermentations.Results
The Streptomyces coelicolor TAG biosynthesis pathway, defined by the acyl-CoA:diacylglycerol acyltransferase (DGAT) Sco0958 and the phosphatidic acid phosphatase (PAP) Lppβ, was successfully reconstructed in an E. coli diacylglycerol kinase (dgkA) mutant strain. TAG production in this genetic background was optimized by increasing the levels of the TAG precursors, diacylglycerol and long-chain acyl-CoAs. For this we carried out a series of stepwise optimizations of the chassis by 1) fine-tuning the expression of the heterologous SCO0958 and lpp β genes, 2) overexpression of the S. coelicolor acetyl-CoA carboxylase complex, and 3) mutation of fadE, the gene encoding for the acyl-CoA dehydrogenase that catalyzes the first step of the β-oxidation cycle in E. coli. The best producing strain, MPS13/pET28-0958-ACC/pBAD-LPPβ rendered a cellular content of 4.85% cell dry weight (CDW) TAG in batch cultivation. Process optimization of fed-batch fermentation in a 1-L stirred-tank bioreactor resulted in cultures with an OD600nm of 80 and a product titer of 722.1 mg TAG L-1 at the end of the process.Conclusions
This study represents the highest reported fed-batch productivity of TAG reached by a model non-oleaginous bacterium. The organism used as a platform was an E. coli BL21 derivative strain containing a deletion in the dgkA gene and containing the TAG biosynthesis genes from S. coelicolor. The genetic studies carried out with this strain indicate that diacylglycerol (DAG) availability appears to be one of the main limiting factors to achieve higher yields of the storage compound. Therefore, in order to develop a competitive process for neutral lipid production in E. coli, it is still necessary to better understand the native regulation of the carbon flow metabolism of this organism, and in particular, to improve the levels of DAG biosynthesis.127.
Sánchez-Rodríguez MA Santiago E Arronte-Rosales A Vargas-Guadarrama LA Mendoza-Núñez VM 《Life sciences》2006,78(15):1682-1687
Psychological stress and environmental pollution are frequently associated to urban environment and oxidative stress (OxS). Likewise, OxS is a risk factor for cognitive impairment (CI) in the elderly. Therefore, we hypothesized that the prevalence of CI in subjects of the urban area could be higher than in those of the rural area, and linked to higher OxS. The aim of the study was to determine the relationship between OxS and CI in elderly individuals from rural and urban settings in Mexico. Plasmatic TBARS, plasma total antioxidant status, and the antioxidant enzymes superoxide dismutase (SOD) and glutathione peroxidase (GPx) were measured in 104 urban and 85 rural elderly individuals. Cognitive functions were evaluated through the Mini Mental State Examination. We found a greater proportion of subjects with OxS and CI in urban than in rural areas (25% vs. 9%), with an odds ratio of 5.67 (CI95% 1.14-38.02, p < 0.05). Our findings suggest that the elderly in urban areas have more OxS and a higher risk of developing CI compared with elderly individuals in a rural environment. 相似文献
128.
Miguel de Mulder Gonzalo Yebra Adriana Navas María Isabel de José María Dolores Gurbindo María Isabel González-Tomé María José Mellado Jesús Saavedra-Lozano María ángeles Mu?oz-Fernández Santiago Jiménez de Ory José Tomás Ramos áfrica Holguín Madrid Cohort of HIV-Infected Children 《PloS one》2012,7(12)
129.
PARP-1 Regulates Metastatic Melanoma through Modulation of Vimentin-induced Malignant Transformation
María Isabel Rodríguez Andreína Peralta-Leal Francisco O'Valle José Manuel Rodriguez-Vargas Ariannys Gonzalez-Flores Jara Majuelos-Melguizo Laura López Santiago Serrano Antonio García de Herreros Juan Carlos Rodríguez-Manzaneque Rubén Fernández Raimundo G. del Moral José Mariano de Almodóvar F. Javier Oliver 《PLoS genetics》2013,9(6)
PARP inhibition can induce anti-neoplastic effects when used as monotherapy or in combination with chemo- or radiotherapy in various tumor settings; however, the basis for the anti-metastasic activities resulting from PARP inhibition remains unknown. PARP inhibitors may also act as modulators of tumor angiogenesis. Proteomic analysis of endothelial cells revealed that vimentin, an intermediary filament involved in angiogenesis and a specific hallmark of EndoMT (endothelial to mesenchymal transition) transformation, was down-regulated following loss of PARP-1 function in endothelial cells. VE-cadherin, an endothelial marker of vascular normalization, was up-regulated in HUVEC treated with PARP inhibitors or following PARP-1 silencing; vimentin over-expression was sufficient to drive to an EndoMT phenotype. In melanoma cells, PARP inhibition reduced pro-metastatic markers, including vasculogenic mimicry. We also demonstrated that vimentin expression was sufficient to induce increased mesenchymal/pro-metastasic phenotypic changes in melanoma cells, including ILK/GSK3-β-dependent E-cadherin down-regulation, Snail1 activation and increased cell motility and migration. In a murine model of metastatic melanoma, PARP inhibition counteracted the ability of melanoma cells to metastasize to the lung. These results suggest that inhibition of PARP interferes with key metastasis-promoting processes, leading to suppression of invasion and colonization of distal organs by aggressive metastatic cells. 相似文献
130.