全文获取类型
收费全文 | 5029篇 |
免费 | 503篇 |
出版年
2023年 | 28篇 |
2022年 | 84篇 |
2021年 | 133篇 |
2020年 | 120篇 |
2019年 | 135篇 |
2018年 | 156篇 |
2017年 | 184篇 |
2016年 | 214篇 |
2015年 | 271篇 |
2014年 | 336篇 |
2013年 | 367篇 |
2012年 | 425篇 |
2011年 | 373篇 |
2010年 | 213篇 |
2009年 | 204篇 |
2008年 | 248篇 |
2007年 | 272篇 |
2006年 | 233篇 |
2005年 | 201篇 |
2004年 | 167篇 |
2003年 | 167篇 |
2002年 | 145篇 |
2001年 | 91篇 |
2000年 | 79篇 |
1999年 | 75篇 |
1998年 | 47篇 |
1997年 | 38篇 |
1996年 | 31篇 |
1995年 | 32篇 |
1994年 | 33篇 |
1993年 | 27篇 |
1992年 | 29篇 |
1991年 | 30篇 |
1990年 | 22篇 |
1989年 | 38篇 |
1988年 | 16篇 |
1987年 | 19篇 |
1986年 | 25篇 |
1985年 | 18篇 |
1984年 | 15篇 |
1983年 | 13篇 |
1982年 | 12篇 |
1979年 | 14篇 |
1978年 | 12篇 |
1977年 | 16篇 |
1976年 | 14篇 |
1975年 | 14篇 |
1974年 | 11篇 |
1972年 | 13篇 |
1966年 | 12篇 |
排序方式: 共有5532条查询结果,搜索用时 15 毫秒
881.
882.
Teresa Almeida Isabel Alonso Sandra Martins Eliana Marisa Ramos Luísa Azevedo Kinji Ohno António Amorim Maria Luiza Saraiva-Pereira Laura Bannach Jardim Tohru Matsuura Jorge Sequeiros Isabel Silveira 《PloS one》2009,4(2)
Spinocerebellar ataxia type 10 (SCA10) is an autosomal dominant neurodegenerative disease characterized by cerebellar ataxia and seizures. The disease is caused by a large ATTCT repeat expansion in the ATXN10 gene. The first families reported with SCA10 were of Mexican origin, but the disease was soon after described in Brazilian families of mixed Portuguese and Amerindian ancestry. The origin of the SCA10 expansion and a possible founder effect that would account for its geographical distribution have been the source of speculation over the last years. To unravel the mutational origin and spread of the SCA10 expansion, we performed an extensive haplotype study, using closely linked STR markers and intragenic SNPs, in families from Brazil and Mexico. Our results showed (1) a shared disease haplotype for all Brazilian and one of the Mexican families, and (2) closely-related haplotypes for the additional SCA10 Mexican families; (3) little or null genetic distance in small normal alleles of different repeat sizes, from the same SNP lineage, indicating that they are being originated by a single step mechanism; and (4) a shared haplotype for pure and interrupted expanded alleles, pointing to a gene conversion model for its generation. In conclusion, we show evidence for an ancestral common origin for SCA10 in Latin America, which might have arisen in an ancestral Amerindian population and later have been spread into the mixed populations of Mexico and Brazil. 相似文献
883.
Dolores Pérez-Sala Patricia Boya Irene Ramos Mónica Herrera Konstantinos Stamatakis 《PloS one》2009,4(12)
Background
Protein degradation is essential for cell homeostasis. Targeting of proteins for degradation is often achieved by specific protein sequences or posttranslational modifications such as ubiquitination.Methodology/Principal Findings
By using biochemical and genetic tools we have monitored the localization and degradation of endogenous and chimeric proteins in live primary cells by confocal microscopy and ultra-structural analysis. Here we identify an eight amino acid sequence from the C-terminus of the short-lived GTPase RhoB that directs the rapid degradation of both RhoB and chimeric proteins bearing this sequence through a lysosomal pathway. Elucidation of the RhoB degradation pathway unveils a mechanism dependent on protein isoprenylation and palmitoylation that involves sorting of the protein into multivesicular bodies, mediated by the ESCRT machinery. Moreover, RhoB sorting is regulated by late endosome specific lipid dynamics and is altered in human genetic lipid traffic disease.Conclusions/Significance
Our findings characterize a short-lived cytosolic protein that is degraded through a lysosomal pathway. In addition, we define a novel motif for protein sorting and rapid degradation, which allows controlling protein levels by means of clinically used drugs. 相似文献884.
Kenneth S. Ramos Adrian Nanez 《Birth defects research. Part C, Embryo today : reviews》2009,87(2):192-197
Recent studies have identified AHR as a master regulator of Wilms' tumor suppressor gene (WT1) signaling in the developing kidney. Activation of AHR signaling by environmental chemical is associated with proteasome‐mediated degradation of AHR protein, disruption of WT1 alternative splicing, and marked alterations in the regulation of genetic programs of developmental progression in the developing kidney. The complexity of genetic regulatory networks of nephrogenesis controlled by AHR‐WT1 interactions will be discussed here with particular emphasis given to the biological and medical consequences that may result from deficits in nephrogenesis that compromise reserve capacity and renal function later in life. Understanding the impact of early‐life environmental exposures to chemicals that disrupt AHR signaling can help minimize negative health consequences to pregnant women and their offspring. Birth Defects Research (Part C) 87:192–197, 2009. © 2009 Wiley‐Liss, Inc. 相似文献
885.
Anésia A. Santos Claudine M. Carvalho Lilian H. Florentino Humberto J. O. Ramos Elizabeth P. B. Fontes 《PloS one》2009,4(6)
NSP-interacting kinase (NIK1) is a receptor-like kinase identified as a virulence target of the begomovirus nuclear shuttle protein (NSP). We found that NIK1 undergoes a stepwise pattern of phosphorylation within its activation-loop domain (A-loop) with distinct roles for different threonine residues. Mutations at Thr-474 or Thr-468 impaired autophosphorylation and were defective for kinase activation. In contrast, a mutation at Thr-469 did not impact autophosphorylation and increased substrate phosphorylation, suggesting an inhibitory role for Thr-469 in kinase function. To dissect the functional significance of these results, we used NSP-expressing virus infection as a mechanism to interfere with wild type and mutant NIK1 action in plants. The NIK1 knockout mutant shows enhanced susceptibility to virus infections, a phenotype that could be complemented with ectopic expression of a 35S-NIK1 or 35S-T469A NIK1 transgenes. However, ectopic expression of an inactive kinase or the 35S-T474A NIK1 mutant did not reverse the enhanced susceptibility phenotype of knockout lines, demonstrating that Thr-474 autophosphorylation was needed to transduce a defense response to geminiviruses. Furthermore, mutations at Thr-474 and Thr-469 residues antagonistically affected NIK-mediated nuclear relocation of the downstream effector rpL10. These results establish that NIK1 functions as an authentic defense receptor as it requires activation to elicit a defense response. Our data also suggest a model whereby phosphorylation-dependent activation of a plant receptor-like kinase enables the A-loop to control differentially auto- and substrate phosphorylation. 相似文献
886.
Aline Machado de Castro Marcelle Lins de Albuquerque de Carvalho Selma Gomes Ferreira Leite Nei Pereira Jr. 《Journal of industrial microbiology & biotechnology》2010,37(2):151-158
The objective of this work is to investigate the utilization of two abundant agricultural residues in Brazil for the production and application of cellulolytic enzymes. Different materials obtained after pretreatment of sugarcane bagasse, as well as pure synthetic substrates, were considered for cellulase production by Penicillium funiculosum. The best results for FPase (354 U L?1) and β-glucosidase (1,835 U L?1) production were observed when sugarcane bagasse partially delignified cellulignin (PDC) was used. The crude extract obtained from PDC fermentation was then partially characterized. Optimal temperatures for cellulase action ranged from 52 to 58°C and pH values of around 4.9 contributed to maximum enzyme activity. At 37°C, the cellulases were highly stable, losing less than 15% of their initial activity after 23 h of incubation. There was no detection of proteases in the P. funiculosum extract, but other hydrolases, such as endoxylanases, were identified (147 U L?1). Finally, when compared to commercial preparations, the cellulolytic complex from P. funiculosum showed more well-balanced amounts of β-glucosidase, endo- and exoglucanase, resulting in the desired performance in the presence of a lignocellulosic material. Cellulases from this filamentous fungus had a higher glucose production rate (470 mg L?1 h?1) when incubated with corn cob than with Celluclast®, GC 220® and Spezyme® (312, 454 and 400 mg L?1 h?1, respectively). 相似文献
887.
There are many different kinds of frameworks for evaluating environmental and sustainability performance at the organizational level (profit or not-for-profit, private or public), sectoral level (e.g. industry, transport, agriculture and tourism), and local, regional or country levels. Despite the diversity of methods and tools to measure sustainable development, indicators are one of the approaches most used. However, these tools do not usually include evaluation of the performance measurement instrument itself. The main objective of this research is to develop a conceptual framework to design and assess the effectiveness of the sustainability indicators themselves. To put the proposed tool into practice, a set of key good-practice factors and meta-performance evaluation indicators is proposed for adoption in a national case study—the national sustainable development indicators system, SIDS Portugal, and the usefulness of this methodology is demonstrated. This approach aims to evaluate how appropriate a set of sustainability indicators is and allow an evaluation of overall performance-monitoring activities and results. Stakeholder involvement is an essential component of the proposed framework. The tool developed could support continuous improvement in the performance of ongoing sustainability indicator initiatives, allowing greater guidance, objectivity and transparency in sustainability assessment processes. 相似文献
888.
Maynes JT Luu HA Cherney MM Andersen RJ Williams D Holmes CF James MN 《Journal of molecular biology》2006,356(1):111-120
The microcystins and nodularins are tumour promoting hepatotoxins that are responsible for global adverse human health effects and wildlife fatalities in countries where drinking water supplies contain cyanobacteria. The toxins function by inhibiting broad specificity Ser/Thr protein phosphatases in the host cells, thereby disrupting signal transduction pathways. A previous crystal structure of a microcystin bound to the catalytic subunit of protein phosphatase-1 (PP-1c) showed distinct changes in the active site region when compared with protein phosphatase-1 structures bound to other toxins. We have elucidated the crystal structures of the cyanotoxins, motuporin (nodularin-V) and dihydromicrocystin-LA bound to human protein phosphatase-1c (gamma isoform). The atomic structures of these complexes reveal the structural basis for inhibition of protein phosphatases by these toxins. Comparisons of the structures of the cyanobacterial toxin:phosphatase complexes explain the biochemical mechanism by which microcystins but not nodularins permanently modify their protein phosphatase targets by covalent addition to an active site cysteine residue. 相似文献
889.
Non-nucleoside inhibitors binding to hepatitis C virus NS5B polymerase reveal a novel mechanism of inhibition 总被引:3,自引:0,他引:3
Biswal BK Wang M Cherney MM Chan L Yannopoulos CG Bilimoria D Bedard J James MN 《Journal of molecular biology》2006,361(1):33-45
The RNA-dependent RNA polymerase (NS5B) from hepatitis C virus (HCV) is a key enzyme in HCV replication. NS5B is a major target for the development of antiviral compounds directed against HCV. Here we present the structures of three thiophene-based non-nucleoside inhibitors (NNIs) bound non-covalently to NS5B. Each of the inhibitors binds to NS5B non-competitively to a common binding site in the "thumb" domain that is approximately 35 Angstroms from the polymerase active site located in the "palm" domain. The three compounds exhibit IC(50) values in the range of 270 nM to 307 nM and have common binding features that result in relatively large conformational changes of residues that interact directly with the inhibitors as well as for other residues adjacent to the binding site. Detailed comparisons of the unbound NS5B structure with those having the bound inhibitors present show that residues Pro495 to Arg505 (the N terminus of the "T" helix) exhibit some of the largest changes. It has been reported that Pro495, Pro496, Val499 and Arg503 are part of the guanosine triphosphate (GTP) specific allosteric binding site located in close proximity to our binding site. It has also been reported that the introduction of mutations to key residues in this region (i.e. Val499Gly) ablate in vivo sub-genomic HCV RNA replication. The details of NS5B polymerase/inhibitor binding interactions coupled with the observed induced conformational changes provide new insights into the design of novel NNIs of HCV. 相似文献
890.
Maia LF Soares MR Valente AP Almeida FC Oliveira AC Gomes AM Freitas MS Schneemann A Johnson JE Silva JL 《The Journal of biological chemistry》2006,281(39):29278-29286
The gamma(1)-peptide is a 21-residue lipid-binding domain from the non-enveloped Flock House virus (FHV). Unlike enveloped viruses, the entry of non-enveloped viruses into cells is believed to occur without membrane fusion. In this study, we performed NMR experiments to establish the solution structure of a membrane-binding peptide from a small non-enveloped icosahedral virus. The three-dimensional structure of the FHV gamma(1)-domain was determined at pH 6.5 and 4.0 in a hydrophobic environment. The secondary and tertiary structures were evaluated in the context of the capacity of the peptide for permeabilizing membrane vesicles of different lipid composition, as measured by fluorescence assays. At both pH values, the peptide has a kinked structure, similar to the fusion domain from the enveloped viruses. The secondary structure was similar in three different hydrophobic environments as follows: water/trifluoroethanol, SDS, and membrane vesicles of different compositions. The ability of the peptide to induce vesicle leakage was highly dependent on the membrane composition. Although the gamma-peptide shares some structural properties to fusion domains of enveloped viruses, it did not induce membrane fusion. Our results suggest that small protein components such as the gamma-peptide in nodaviruses (such as FHV) and VP4 in picornaviruses have a crucial role in conducting nucleic acids through cellular membranes and that their structures resemble the fusion domains of membrane proteins from enveloped viruses. 相似文献