全文获取类型
收费全文 | 5927篇 |
免费 | 461篇 |
国内免费 | 2篇 |
出版年
2024年 | 5篇 |
2023年 | 18篇 |
2022年 | 62篇 |
2021年 | 123篇 |
2020年 | 88篇 |
2019年 | 99篇 |
2018年 | 176篇 |
2017年 | 147篇 |
2016年 | 234篇 |
2015年 | 336篇 |
2014年 | 407篇 |
2013年 | 461篇 |
2012年 | 556篇 |
2011年 | 529篇 |
2010年 | 346篇 |
2009年 | 294篇 |
2008年 | 377篇 |
2007年 | 331篇 |
2006年 | 310篇 |
2005年 | 266篇 |
2004年 | 289篇 |
2003年 | 194篇 |
2002年 | 179篇 |
2001年 | 135篇 |
2000年 | 91篇 |
1999年 | 76篇 |
1998年 | 32篇 |
1997年 | 34篇 |
1996年 | 23篇 |
1995年 | 17篇 |
1994年 | 11篇 |
1993年 | 14篇 |
1992年 | 21篇 |
1991年 | 18篇 |
1990年 | 22篇 |
1989年 | 15篇 |
1988年 | 7篇 |
1987年 | 9篇 |
1986年 | 3篇 |
1985年 | 8篇 |
1984年 | 2篇 |
1983年 | 2篇 |
1982年 | 3篇 |
1979年 | 2篇 |
1978年 | 3篇 |
1977年 | 3篇 |
1975年 | 2篇 |
1974年 | 3篇 |
1967年 | 2篇 |
1961年 | 1篇 |
排序方式: 共有6390条查询结果,搜索用时 359 毫秒
931.
932.
Soyoung Park Young‐Ha Park Chang‐Ro Lee Yeon‐Ran Kim Yeong‐Jae Seok 《Molecular microbiology》2016,101(5):795-808
To survive in a continuously changing environment, bacteria sense concentration gradients of attractants or repellents, and purposefully migrate until a more favourable habitat is encountered. While glucose is known as the most effective attractant, the flagellar biosynthesis and hence chemotactic motility has been known to be repressed by glucose in some bacteria. To date, the only known regulatory mechanism of the repression of flagellar synthesis by glucose is via downregulation of the cAMP level, as shown in a few members of the family Enterobacteriaceae. Here we show that, in Vibrio vulnificus, the glucose‐mediated inhibition of flagellar motility operates by a completely different mechanism. In the presence of glucose, EIIAGlc is dephosphorylated and inhibits the polar localization of FapA (flagellar assembly protein A) by sequestering it from the flagellated pole. A loss or delocalization of FapA results in a complete failure of the flagellar biosynthesis and motility. However, when glucose is depleted, EIIAGlc is phosphorylated and releases FapA such that free FapA can be localized back to the pole and trigger flagellation. Together, these data provide new insight into a bacterial strategy to reach and stay in the glucose‐rich area. 相似文献
933.
As healthcare organizations are struggling to enhance healthcare qualities and efficiencies, they focus on secure and efficient sharing of clinical information among healthcare providers including doctors, nurses, medical laboratories, and informal caregivers, throughout the entire healthcare life cycle. Any failure or error during this clinical collaborative communication can have negative impact on the efficiencies of the whole healthcare delivery system. This paper suggests a methodological approach to address the needs in this area and proposes a methodology for developing clinical collaborative communication (C\(^{3})\) systems. Since C\(^{3}\) demands a variety of collaborative communications among multiple healthcare stakeholders, the proposed approach focuses on the effective system design and analysis with the objective to enhance collaboration and communication in healthcare organizations. 相似文献
934.
The Escherichia coli proteome response to the stressor GdnHCl was analyzed through 2-dimensional gel electrophoresis (2-DE). We identified PotD (spermidine/putrescine-binding periplasmic protein) and Crr [glucose-specific phosphotransferase (PTS) enzyme IIA component] as a stress-responsive protein. Even under a stress situation where the total number of soluble proteins decreased by about 10%, 3.5- and 2.2-fold increase was observed in the synthesis of PotD and Crr, respectively. As fusion partners, PotD and Crr dramatically increased the solubility of many aggregation-prone heterologous proteins [e.g. human minipro-insulin (mp-INS), human epidermal growth factor (EGF), human prepro-ghrelin (ppGRN), human interleukin-2(hIL-2), human activation induced cytidine deaminase (AID), human glutamate decarboxylase (GAD(448-585)), Pseudomonas putida cutinase (CUT), human ferritin light chain (hFTN-L), human granulocyte colony-stimulating factor (G-CSF), and cold autoinflammatory syndrome1 protein (NALP3) Nacht domain (NACHT)] in the E. coli cytoplasm. Presumably PotD and Crr were very effective in shielding interactive surfaces of heterologous proteins associated with non-specific protein-protein interactions leading to the formation of inclusion bodies most likely due to intrinsic high folding efficiency, chaperone-like activity, or a combination of both factors. Both the stress-induced proteins were well suited for the production of a biologically active fusion mutant of P. putida cutinase that can be expected to be of biotechnological and commercial interest. 相似文献
935.
Novel mechanistic class of fatty acid amide hydrolase inhibitors with remarkable selectivity 总被引:5,自引:0,他引:5
Ahn K Johnson DS Fitzgerald LR Liimatta M Arendse A Stevenson T Lund ET Nugent RA Nomanbhoy TK Alexander JP Cravatt BF 《Biochemistry》2007,46(45):13019-13030
Fatty acid amide hydrolase (FAAH) is an integral membrane enzyme that degrades the fatty acid amide family of signaling lipids, including the endocannabinoid anandamide. Genetic or pharmacological inactivation of FAAH leads to analgesic, anti-inflammatory, anxiolytic, and antidepressant phenotypes in rodents without showing the undesirable side effects observed with direct cannabinoid receptor agonists, indicating that FAAH may represent an attractive therapeutic target for treatment of pain, inflammation, and other central nervous system disorders. However, the FAAH inhibitors reported to date lack drug-like pharmacokinetic properties and/or selectivity. Herein we describe piperidine/piperazine ureas represented by N-phenyl-4-(quinolin-3-ylmethyl)piperidine-1-carboxamide (PF-750) and N-phenyl-4-(quinolin-2-ylmethyl)piperazine-1-carboxamide (PF-622) as a novel mechanistic class of FAAH inhibitors. PF-750 and PF-622 show higher in vitro potencies than previously established classes of FAAH inhibitors. Rather unexpectedly based on the high chemical stability of the urea functional group, PF-750 and PF-622 were found to inhibit FAAH in a time-dependent manner by covalently modifying the enzyme's active site serine nucleophile. Activity-based proteomic profiling revealed that PF-750 and PF-622 were completely selective for FAAH relative to other mammalian serine hydrolases. We hypothesize that this remarkable specificity derives, at least in part, from FAAH's special ability to function as a C(O)-N bond hydrolase, which distinguishes it from the vast majority of metabolic serine hydrolases in mammals that are restricted to hydrolyzing esters and/or thioesters. The piperidine/piperazine urea may thus represent a privileged chemical scaffold for the synthesis of FAAH inhibitors that display an unprecedented combination of potency and selectivity for use as potential analgesic and anxiolytic/antidepressant agents. 相似文献
936.
937.
Katheryn A. Resing Natalie G. Ahn 《Progress in biophysics and molecular biology》1999,71(3-4):501-523
Advances in mass spectrometry instrumentation, protocols for sample handling, and computational methods provide powerful new approaches to solving problems in analytical biochemistry. This review summarizes recent work illustrating ways in which mass spectrometry has been used to address questions relevant to signal transduction. Rather than encompass all of the instruments or methodologies that might be brought to bear on these problems, we present an overview of commonly used techniques, promising new methodologies, and some applications. 相似文献
938.
J Klein-Seetharaman J Hwa K Cai C Altenbach W L Hubbell H G Khorana 《Biochemistry》1999,38(25):7938-7944
Cysteines were introduced, one at a time, at amino acid positions 55-75 in the cytoplasmic region connecting helices I and II in rhodopsin. In each of the 21 cysteine mutants, the reactive native cysteine residues (C140 and C316) were replaced by serine. Except for N55C, all mutants formed rhodopsin-like chromophores and had normal photobleaching characteristics. The efficiency of GT activation was reduced only for K66C, K67C, L68C, and P71C. The reactivity of the substituted cysteine in each mutant toward 4, 4'-dithiodipyridine (4-PDS) was investigated in the dark. The mutants F56C to L59C and I75C were unreactive to 4-PDS under the conditions used, suggesting that they are embedded in the micelle or protein interior. The mutants V63C, H65C-T70C, and N73C reacted rapidly, while the remainder of the mutants reacted more slowly, and varied in reactivity with sequence position. For the mutants derivatized with 4-PDS, the rate of release of thiopyridone from the resulting thiopyridinyl-cysteine disulfide bond by dithiothreitol was investigated in the dark and in the light. Marked changes in the rates of thiopyridone release in the light were found at specific sites. Collectively, the data reveal tertiary interactions of the residues in the sequence investigated and demonstrate structural changes due to photoactivation. 相似文献
939.
Twenty-one single-cysteine substitution mutants were prepared in the sequence 56-75 between transmembrane helices I and II at the cytoplasmic surface of bovine rhodopsin. Each mutant was reacted with a sulfhydryl-specific reagent to produce a nitroxide side chain. The electron paramagnetic resonance of the labeled proteins in dodecyl maltoside solution was analyzed to provide the relative mobility and accessibility of the nitroxide side chain to both polar and nonpolar paramagnetic reagents. The results indicate that the hydrophobic-water interface of the micelle intersects helices I and II near residues 64 and 71, respectively. Thus, the sequence 64-71 is in the aqueous phase, while 56-63 and 72-75 lie in the transmembrane helices I and II, respectively. The lipid-facing surfaces on transmembrane helices I and II near the cytoplasmic surface correspond to approximately 180 degrees and 90 degrees of arc on the helical surfaces, respectively. Photoactivation of rhodopsin produced changes in structure in the region investigated, primarily around helix II. However, these changes are much smaller than those noted by spin labels in helix VI (Altenbach, C., Yang, K., Farrens, D., Farahbakhsh, Z., Khorana, H. G., and Hubbell, W. L. (1996) Biochemistry 35, 12470). 相似文献
940.
S Y Choi F Kwok J H Bahn S G Jeon Y K Ahn B H Yoon B R Lee K S Choi G Z Gao 《BioFactors (Oxford, England)》1999,10(1):35-42
Six monoclonal antibodies that recognize porcine brain pyridoxal kinase have been selected and designated as PK67, PK86, PK91, PK144, PK252 and PK275. A total of six monoclonal antibodies recognizing different epitopes of the enzyme were obtained, of which four inhibited the enzyme activity. When total proteins of porcine brain homogenate separated by SDS-PAGE were subjected to monoclonal antibodies, a single reactive protein band of molecular weight 39 kDa which comigrated with purified porcine pyridoxal kinase was detected. Using the anti-pyridoxal kinase antibodies as probes, the cross reactivities of brain pyridoxal kinase from human and other mammalian tissues and from avian sources were also investigated. Among human and all animal tissues tested, immunoreactive bands on Western blots appeared to have the same molecular mass of 39 kDa. These results indicate that mammalian brains contain only one major type of immunologically similar pyridoxal kinase, although some properties of the enzymes reported previously differed from one another. 相似文献