全文获取类型
收费全文 | 1399篇 |
免费 | 163篇 |
出版年
2023年 | 8篇 |
2022年 | 15篇 |
2021年 | 34篇 |
2020年 | 24篇 |
2019年 | 29篇 |
2018年 | 30篇 |
2017年 | 25篇 |
2016年 | 55篇 |
2015年 | 73篇 |
2014年 | 84篇 |
2013年 | 107篇 |
2012年 | 118篇 |
2011年 | 101篇 |
2010年 | 71篇 |
2009年 | 53篇 |
2008年 | 81篇 |
2007年 | 74篇 |
2006年 | 60篇 |
2005年 | 61篇 |
2004年 | 56篇 |
2003年 | 61篇 |
2002年 | 56篇 |
2001年 | 20篇 |
2000年 | 20篇 |
1999年 | 20篇 |
1998年 | 18篇 |
1997年 | 19篇 |
1996年 | 8篇 |
1995年 | 7篇 |
1994年 | 6篇 |
1993年 | 8篇 |
1992年 | 14篇 |
1991年 | 7篇 |
1990年 | 15篇 |
1989年 | 7篇 |
1988年 | 4篇 |
1987年 | 7篇 |
1986年 | 4篇 |
1985年 | 4篇 |
1984年 | 7篇 |
1983年 | 8篇 |
1982年 | 7篇 |
1981年 | 5篇 |
1980年 | 5篇 |
1979年 | 9篇 |
1977年 | 10篇 |
1976年 | 4篇 |
1967年 | 3篇 |
1932年 | 3篇 |
1929年 | 3篇 |
排序方式: 共有1562条查询结果,搜索用时 31 毫秒
91.
Wissinger B Gamer D Jägle H Giorda R Marx T Mayer S Tippmann S Broghammer M Jurklies B Rosenberg T Jacobson SG Sener EC Tatlipinar S Hoyng CB Castellan C Bitoun P Andreasson S Rudolph G Kellner U Lorenz B Wolff G Verellen-Dumoulin C Schwartz M Cremers FP Apfelstedt-Sylla E Zrenner E Salati R Sharpe LT Kohl S 《American journal of human genetics》2001,69(4):722-737
We recently showed that mutations in the CNGA3 gene encoding the alpha-subunit of the cone photoreceptor cGMP-gated channel cause autosomal recessive complete achromatopsia linked to chromosome 2q11. We now report the results of a first comprehensive screening for CNGA3 mutations in a cohort of 258 additional independent families with hereditary cone photoreceptor disorders. CNGA3 mutations were detected not only in patients with the complete form of achromatopsia but also in incomplete achromats with residual cone photoreceptor function and (rarely) in patients with evidence for severe progressive cone dystrophy. In total, mutations were identified in 53 independent families comprising 38 new CNGA3 mutations, in addition to the 8 mutations reported elsewhere. Apparently, both mutant alleles were identified in 47 families, including 16 families with presumed homozygous mutations and 31 families with two heterozygous mutations. Single heterozygous mutations were identified in six additional families. The majority of all known CNGA3 mutations (39/46) are amino acid substitutions compared with only four stop-codon mutations, two 1-bp insertions and one 3-bp in-frame deletion. The missense mutations mostly affect amino acids conserved among the members of the cyclic nucleotide gated (CNG) channel family and cluster at the cytoplasmic face of transmembrane domains (TM) S1 and S2, in TM S4, and in the cGMP-binding domain. Several mutations were identified recurrently (e.g., R277C, R283W, R436W, and F547L). These four mutations account for 41.8% of all detected mutant CNGA3 alleles. Haplotype analysis suggests that the R436W and F547L mutant alleles have multiple origins, whereas we found evidence that the R283W alleles, which are particularly frequent among patients from Scandinavia and northern Italy, have a common origin. 相似文献
92.
The carboxy-terminal acidic domain of Rift Valley Fever virus NSs protein is essential for the formation of filamentous structures but not for the nuclear localization of the protein 下载免费PDF全文
The ambisense S segment of Rift Valley fever (RVF) virus (a phlebovirus in the Bunyaviridae family) codes for two proteins: the viral complementary-sense RNA for the N nucleoprotein and the genomic-sense RNA for the nonstructural protein NSs. Except for the fact that the NSs protein is phosphorylated and forms filamentous structures in the nuclei of infected cells (R. Swanepoel and N. K. Blackburn, J. Gen. Virol. 34:557-561, 1977), its role is poorly understood, especially since the replication cycle of all these viruses takes place in the cytoplasm. To investigate the mechanisms involved in filament formation, we expressed NSs in mammalian cells via a recombinant Semliki Forest virus and demonstrated that the protein alone was able to form structures similar to those observed in RVF virus-infected cells, indicating that the presence of other RVF virus proteins is not required for filament formation. The yeast two-hybrid system was used to show that the protein interacts with itself and to map the interacting domains. Various deletion and substitution mutants were constructed, and the mutant proteins were analyzed by immunoprecipitation, Western blotting and immunofluorescence. These experiments indicated that the 10 to 17 amino acids of the carboxy-terminal domain were involved in self-association of the protein and that deletion of this acidic carboxy-terminal domain prevents the protein from forming filaments but does not affect its nuclear localization. The role of two phosphorylation sites present in this domain was also investigated, but they were not found to have a major influence on the formation of the nuclear filament. 相似文献
93.
Clinical and research data indicate that active and passive changes in the mechanical environment of the heart are capable of influencing both the initiation and the spread of cardiac excitation via pathways that are intrinsic to the heart. This direction of the cross-talk between cardiac electrical and mechanical activity is referred to as mechano-electric feedback (MEF). MEF is thought to be involved in the adjustment of heart rate to changes in mechanical load and would help to explain the precise beat-to-beat regulation of cardiac performance as it occurs even in the recently transplanted (and, thus, denervated) heart. Furthermore, there is clinical evidence that MEF may be involved in mechanical initiation of arrhythmias and fibrillation, as well as in the re-setting of disturbed heart rhythm by 'mechanical' first aid procedures. This review will outline the clinical relevance of cardiac MEF, describe cellular correlates to the responses observed in situ, and discuss the role that quantitative mathematical models may play in identifying the involvement of cardiac MEF in the regulation of heart rate and rhythm. 相似文献
94.
In the human demyelinating disorder multiple sclerosis, and its animal model experimental allergic encephalomyelitis, there is a breakdown of the blood-brain barrier and an infiltration of immune cells into the CNS. Infiltrating T lymphocytes and macrophages are believed to be key mediators of the disease process. Considerable circumstantial and experimental evidence has suggested that the pleiotropic cytokine interferon gamma (IFN-), which is exclusively expressed by T cells and natural killer cells, is a deleterious component of the immune response in these disorders. When experimentally introduced into the CNS IFN- promotes many of the pathological changes that occur in immune-mediated demyelinating disorders. In vitro, this cytokine elicits a number of effects on oligodendrocytes, including cell death. The harmful actions of IFN- on CNS myelin are likely mediated through direct effects on the myelinating cells, as well as through the activation of macrophages and microglia. In this review we summarize relevant studies concerning the action of IFN- in demyelinating disorders and discuss possible mechanisms for the observed effects. 相似文献
95.
96.
Binz HK Amstutz P Kohl A Stumpp MT Briand C Forrer P Grütter MG Plückthun A 《Nature biotechnology》2004,22(5):575-582
We report here the evolution of ankyrin repeat (AR) proteins in vitro for specific, high-affinity target binding. Using a consensus design strategy, we generated combinatorial libraries of AR proteins of varying repeat numbers with diversified binding surfaces. Libraries of two and three repeats, flanked by 'capping repeats,' were used in ribosome-display selections against maltose binding protein (MBP) and two eukaryotic kinases. We rapidly enriched target-specific binders with affinities in the low nanomolar range and determined the crystal structure of one of the selected AR proteins in complex with MBP at 2.3 A resolution. The interaction relies on the randomized positions of the designed AR protein and is comparable to natural, heterodimeric protein-protein interactions. Thus, our AR protein libraries are valuable sources for binding molecules and, because of the very favorable biophysical properties of the designed AR proteins, an attractive alternative to antibody libraries. 相似文献
97.
Growth of high quality crystals is often the most difficult step in the determination of protein structures by X-ray diffraction. Automation can improve the success of this process both by reducing the amount of protein required for each screen and by relieving the tedium of setting up crystallization experiments by hand. We have been using an automated system for the design and execution of hanging drop crystallization experiments for the last two years. The system includes robots for the preparation of solutions, setup of hanging drops, and automated imaging, as well as a new software package (RoCKS) for managing all phases of the crystallization process. Here, we review the fundamentals of automated protein crystallization and present results from our comparisons of various approaches to screening. 相似文献
98.
Hiller S Kohl A Fiorito F Herrmann T Wider G Tschopp J Grütter MG Wüthrich K 《Structure (London, England : 1993)》2003,11(10):1199-1205
Signaling in apoptosis and inflammation is often mediated by proteins of the death domain superfamily in the Fas/FADD/Caspase-8 or the Apaf-1/Caspase-9 pathways. This superfamily currently comprises the death domain (DD), death effector domain (DED), caspase recruitment domain (CARD), and pyrin domain (PYD) subfamilies. The PYD subfamily is most abundant, but three-dimensional structures are only available for the subfamilies DD, DED, and CARD, which have an antiparallel arrangement of six alpha helices as common fold. This paper presents the NMR structure of PYD of NALP1, a protein that is involved in the innate immune response and is a component of the inflammasome. The structure of NALP1 PYD differs from all other known death domain superfamily structures in that the third alpha helix is replaced by a flexibly disordered loop. This unique feature appears to relate to the molecular basis of familial Mediterranean fever (FMF), a genetic disease caused by single-point mutations. 相似文献
99.
Helfand MS Bethel CR Hujer AM Hujer KM Anderson VE Bonomo RA 《The Journal of biological chemistry》2003,278(52):52724-52729
Bacterial resistance to beta-lactam/beta-lactamase inhibitor combinations by single amino acid mutations in class A beta-lactamases threatens our most potent clinical antibiotics. In TEM-1 and SHV-1, the common class A beta-lactamases, alterations at Ser-130 confer resistance to inactivation by the beta-lactamase inhibitors, clavulanic acid, and tazobactam. By using site-saturation mutagenesis, we sought to determine the amino acid substitutions at Ser-130 in SHV-1 beta-lactamase that result in resistance to these inhibitors. Antibiotic susceptibility testing revealed that ampicillin and ampicillin/clavulanic acid resistance was observed only for the S130G beta-lactamase expressed in Escherichia coli. Kinetic analysis of the S130G beta-lactamase demonstrated a significant elevation in apparent Km and a reduction in kcat/Km for ampicillin. Marked increases in the dissociation constant for the preacylation complex, KI, of clavulanic acid (SHV-1, 0.14 microm; S130G, 46.5 microm) and tazobactam (SHV-1, 0.07 microm; S130G, 4.2 microm) were observed. In contrast, the k(inact)s of S130G and SHV-1 differed by only 17% for clavulanic acid and 40% for tazobactam. Progressive inactivation studies showed that the inhibitor to enzyme ratios required to inactivate SHV-1 and S130G were similar. Our observations demonstrate that enzymatic activity is preserved despite amino acid substitutions that significantly alter the apparent affinity of the active site for beta-lactams and beta-lactamase inhibitors. These results underscore the mechanistic versatility of class A beta-lactamases and have implications for the design of novel beta-lactamase inhibitors. 相似文献
100.
Human ADAM33: protein maturation and localization 总被引:6,自引:0,他引:6
Garlisi CG Zou J Devito KE Tian F Zhu FX Liu J Shah H Wan Y Motasim Billah M Egan RW Umland SP 《Biochemical and biophysical research communications》2003,301(1):35-43
ADAM33 (a disintegrin and metalloprotease) was recently found to be a novel asthma susceptibility gene. Domain-specific antibodies were used to study its expression and processing. When the pro-domain and catalytic domain were expressed by a stable-transfected cell line, the pro-domain was removed by cleavage within a putative furin cleavage site. The catalytic domain was active in an alpha(2)-macroglobulin complex formation assay and mutation of the catalytic site glutamic acid (E346A) eliminated activity. In transient transfections using the full-length protein, a pro-form and mature form were detectable and alternate glycosylation was demonstrated at sites within the catalytic domain. ADAM33 was detected on the cell surface, with the majority of protein detected intracellularly. The E346A mutation had no significant effect on protein processing. Endogenous ADAM33 was detected in bronchus tissue, bronchial smooth muscle cells, and MRC-5 fibroblasts, consistent with a role in the pathophysiology of asthma. 相似文献