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731.
Hobson RP Munro CA Bates S MacCallum DM Cutler JE Heinsbroek SE Brown GD Odds FC Gow NA 《The Journal of biological chemistry》2004,279(38):39628-39635
The outer layer of the cell wall of the human pathogenic fungus Candida albicans is enriched with heavily mannosylated glycoproteins that are the immediate point of contact between the fungus and cells of the host, including phagocytes. Previous work had identified components of the acid-labile fraction of N-linked mannan, comprising beta-1,2-linked mannose residues attached via a phosphodiester bond, as potential ligands for macrophage receptors and modulators of macrophage function. We therefore isolated and disrupted the CaMNN4 gene, which is required for mannosyl phosphate transfer and hence the attachment of beta-1,2 mannose oligosaccharides to the acid-labile N-mannan side chains. With the mannosylphosphate eliminated, the mnn4Delta null mutant was unable to bind the charged cationic dye Alcian Blue and was devoid of acid-labile beta-1,2-linked oligomannosaccharides. The mnn4Delta mutant was unaffected in cell growth and morphogenesis in vitro and in virulence in a murine model of systemic C. albicans infection. The null mutant was also not affected in its interaction with macrophages. Mannosylphosphate is therefore not required for macrophage interactions or for virulence of C. albicans. 相似文献
732.
Kitzing K Auweter S Amrhein N Macheroux P 《The Journal of biological chemistry》2004,279(10):9451-9461
Chorismate synthase catalyzes the last step in the common shikimate pathway leading to aromatic compounds such as the aromatic amino acids. The reaction consists of the 1,4-anti-elimination of the 3-phosphate group and the C-(6proR) hydrogen from 5-enolpyruvylshikimate 3-phosphate to yield chorismate. Although this reaction does not involve a net redox change, the enzyme has an absolute requirement for reduced flavin mononucleotide, which is not consumed during the reaction. Two invariant histidine residues are found in the active site of the enzyme: His(17) and His(106). Using site-directed mutagenesis, both histidines were replaced by alanine, reducing the activity 10- and 20-fold in the H106A and H17A mutant protein, respectively. Based on the characterization of the two single mutant proteins, it is proposed that His(106) serves to protonate the monoanionic reduced FMN, whereas His(17) protonates the leaving phosphate group of the substrate. An enzymatic reaction mechanism in keeping with the experimental results is presented. 相似文献
733.
Rational engineering of enzyme stability 总被引:11,自引:0,他引:11
Eijsink VG Bjørk A Gåseidnes S Sirevåg R Synstad B van den Burg B Vriend G 《Journal of biotechnology》2004,113(1-3):105-120
During the past 15 years there has been a continuous flow of reports describing proteins stabilized by the introduction of mutations. These reports span a period from pioneering rational design work on small enzymes such as T4 lysozyme and barnase to protein design, and directed evolution. Concomitantly, the purification and characterization of naturally occurring hyperstable proteins has added to our understanding of protein stability. Along the way, many strategies for rational protein stabilization have been proposed, some of which (e.g. entropic stabilization by introduction of prolines or disulfide bridges) have reasonable success rates. On the other hand, comparative studies and efforts in directed evolution have revealed that there are many mutational strategies that lead to high stability, some of which are not easy to define and rationalize. Recent developments in the field include increasing awareness of the importance of the protein surface for stability, as well as the notion that normally a very limited number of mutations can yield a large increase in stability. Another development concerns the notion that there is a fundamental difference between the "laboratory stability" of small pure proteins that unfold reversibly and completely at high temperatures and "industrial stability", which is usually governed by partial unfolding processes followed by some kind of irreversible inactivation process (e.g. aggregation). Provided that one has sufficient knowledge of the mechanism of thermal inactivation, successful and efficient rational stabilization of enzymes can be achieved. 相似文献
734.
Sigrid?B.?Schnack-SchielEmail author Gerhard?S.?Dieckmann Gerhard?Kattner David?N.?Thomas 《Polar Biology》2004,27(8):502-506
Copepods in platelet-ice layers underlying fast ice and in the water column below were studied at Drescher Inlet, eastern Weddell Sea in February 1998. Three copepod species were found: Drescheriella glacialis and Paralabidocera antarctica occurred in platelet-ice layers, while Stephos longipes was only present in the water column. The distribution of all species varied considerably between station and depth. D. glacialis dominated the platelet-ice community and occurred at all five platelet-ice sampling sites, except one, with numbers of up to 26 ind. l–1. In contrast, P. antarctica was only found in low numbers (up to 2 ind. l–1) at one site. The total copepod abundance in the platelet ice was not associated with algal biomass, although it was strongly correlated with high ammonium concentrations (up to 9 M) in the interstitial water between the platelets. This is the first indirect evidence to support the hypothesis that zooplankton excretion can partly account for the high ammonium values often found in platelet-ice layers. 相似文献
735.
Marchal K De Keersmaecker S Monsieurs P van Boxel N Lemmens K Thijs G Vanderleyden J De Moor B 《Genome biology》2004,5(2):R9
Background
The PmrAB (BasSR) two-component regulatory system is required for Salmonella typhimurium virulence. PmrAB-controlled modifications of the lipopolysaccharide (LPS) layer confer resistance to cationic antibiotic polypeptides, which may allow bacteria to survive within macrophages. The PmrAB system also confers resistance to Fe3+-mediated killing. New targets of the system have recently been discovered that seem not to have a role in the well-described functions of PmrAB, suggesting that the PmrAB-dependent regulon might contain additional, unidentified targets. 相似文献736.
737.
738.
Nonredundant Regulation of Rice Arbuscular Mycorrhizal Symbiosis by Two Members of the PHOSPHATE TRANSPORTER1 Gene Family 总被引:1,自引:0,他引:1
Shu-Yi Yang Mette Gr?nlund Iver Jakobsen Marianne Suter Grotemeyer Doris Rentsch Akio Miyao Hirohiko Hirochika Chellian Santhosh Kumar Venkatesan Sundaresan Nicolas Salamin Sheryl Catausan Nicolas Mattes Sigrid Heuer Uta Paszkowski 《The Plant cell》2012,24(10):4236-4251
Pi acquisition of crops via arbuscular mycorrhizal (AM) symbiosis is becoming increasingly important due to limited high-grade rock Pi reserves and a demand for environmentally sustainable agriculture. Here, we show that 70% of the overall Pi acquired by rice (Oryza sativa) is delivered via the symbiotic route. To better understand this pathway, we combined genetic, molecular, and physiological approaches to determine the specific functions of two symbiosis-specific members of the PHOSPHATE TRANSPORTER1 (PHT1) gene family from rice, ORYsa;PHT1;11 (PT11) and ORYsa;PHT1;13 (PT13). The PT11 lineage of proteins from mono- and dicotyledons is most closely related to homologs from the ancient moss, indicating an early evolutionary origin. By contrast, PT13 arose in the Poaceae, suggesting that grasses acquired a particular strategy for the acquisition of symbiotic Pi. Surprisingly, mutations in either PT11 or PT13 affected the development of the symbiosis, demonstrating that both genes are important for AM symbiosis. For symbiotic Pi uptake, however, only PT11 is necessary and sufficient. Consequently, our results demonstrate that mycorrhizal rice depends on the AM symbiosis to satisfy its Pi demands, which is mediated by a single functional Pi transporter, PT11. 相似文献
739.
Maisch T Shimizu T Mitra A Heinlin J Karrer S Li YF Morfill G Zimmermann JL 《Journal of industrial microbiology & biotechnology》2012,39(9):1367-1375
In this study we investigated the sensitivity of Deinococcus radiodurans to contact-free cold atmospheric plasma treatment as part of a project to establish new efficient procedures for disinfection of inanimate surfaces. The Gram-positive D.?radiodurans is one of the most resistant microorganisms worldwide. Stationary phases of D.?radiodurans were exposed to cold atmospheric plasma for different time intervals or to ultraviolet?C (UVC) radiation at dose rates of 0.001-0.0656?J?cm(-2), respectively. A methicillin-resistant Staphylococcus aureus strain (MRSA) served as control for Gram-positive bacteria. The surface microdischarge plasma technology was used for generation of cold atmospheric plasma. A plasma discharge was ignited using ambient air. Surprisingly, D.?radiodurans was sensitive to the cold atmospheric plasma treatment in the same range as the MRSA strain. Survival of both bacteria decreased with increasing plasma exposure times up to 6 log(10) cycles (>99.999?%) within 20?s of plasma treatment. In contrast, UVC radiation of both bacteria demonstrated that D.?radiodurans was more resistant to UVC treatment than MRSA. Cold atmospheric plasma seems to be a promising tool for industrial and clinical purposes where time-saving is a critical point to achieve efficient disinfection of inanimate surfaces and where protection from corrosive materials is needed. 相似文献
740.
van Beveren NJ Buitendijk GH Swagemakers S Krab LC Röder C de Haan L van der Spek P Elgersma Y 《PloS one》2012,7(2):e32618