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1.
The involvement of O-sulphate esters in the directed O-methylation was investigated in vitro with a dialysed "high-speed' supernatant from rat liver as the enzyme preparation and the catechol compound 3,4-dihydroxybenzoic acid as the substrate. The enzyme reactions involved were studied separately with the O-methylated and O-sulphated derivatives. The rate of hydrolysis by arylsulphatase was 14.5 nmol/min per mg of protein for 3-methoxy-4-sulphonyloxybenzoic acid and 10.1 nmol/min per mg of protein for 4-methoxy-3-sulphonyloxybenzoic acid. The sulphotransferase activity towards the guaiacols 4-hydroxy-3-methoxybenzoic acid and 3-hydroxy-4-methoxybenzoic acid was 570pmol of 4-O-sulphated and 350pmol of 3-O-sulphated product formed/min per mg of protein. The 3-O- and 4-O-sulphate esters of 3,4-dihydroxybenzoic acid could not serve as substrates for the catechol O-methyltransferase reaction. When either ester was incubated in the presence of S-adenosyl-L-methionine, but without the arylsulphatase inhibitor KH2PO4, 3,4-dihydroxybenzoic acid was formed, which was subsequently O-methylated in a meta/para ratio of 4.6. It is concluded that O-methylation can precede O-sulphation but that O-sulphation prevents further metabolism by O-methylation. Also O-sulphate esters do not have a directing effect on O-methylation. From the study of the simultaneous action of sulphotransferase and catechol O-methyltransferase on 3,4-dihydroxybenzoic acid we conclude that O-sulphation and O-methylation proceed independently of each other under the assay conditions used, both directed preferentially to the 3-hydroxy group.  相似文献   
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We explore a common feature of insect population dynamics, interspecific synchrony, which refers to synchrony in population dynamics among sympatric populations of different species. Such synchrony can arise via several possible mechanisms, including shared environmental effects and shared trophic interactions, but distinguishing the relative importance among different mechanisms can be challenging. We analyze interannual time series of population densities of the larch budmoth, Zeiraphera griseana (Lepidoptera: Tortricidae), along with six sympatric larch-feeding folivores from a site in the European Alps 1952–1979. These species include five lepidopterans, Exapate duratella, Ptycholomoides aeriferana, Spilonota laricana, Epirrita autumnata and Teleiodes saltuum, and one hymenopteran sawfly Pristiphora laricis. We document that the highly regular oscillatory behavior (period 9–10 years) of Z. griseana populations is similarly evident in the dynamics of most of the sympatric folivores. We also find that all of the sympatric species are phase synchronized with Z. griseana populations with half of the sympatric species exhibiting nonlagged phase synchrony and three of the species exhibiting 2–5 year lags behind Z. griseana populations. We adapt a previously developed tritrophic model of Z. griseana dynamics to explore possible mechanisms responsible for observed phase synchronization. Results suggest that either shared stochastic influences (e.g., weather) or shared parasitoid impacts are likely causes of nonlagged phase synchronization. The model further indicates that observed patterns of lagged phase synchronization are most likely caused by either shared delayed induced host plant defenses or direct density-dependent effects shared with Z. griseana.  相似文献   
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Spinal muscular atrophy (SMA) is a heterogeneous group of neuromuscular disorders caused by degeneration of lower motor neurons. Although functional loss of SMN1 is associated with autosomal-recessive childhood SMA, the genetic cause for most families affected by dominantly inherited SMA is unknown. Here, we identified pathogenic variants in bicaudal D homolog 2 (Drosophila) (BICD2) in three families afflicted with autosomal-dominant SMA. Affected individuals displayed congenital slowly progressive muscle weakness mainly of the lower limbs and congenital contractures. In a large Dutch family, linkage analysis identified a 9q22.3 locus in which exome sequencing uncovered c.320C>T (p.Ser107Leu) in BICD2. Sequencing of 23 additional families affected by dominant SMA led to the identification of pathogenic variants in one family from Canada (c.2108C>T [p.Thr703Met]) and one from the Netherlands (c.563A>C [p.Asn188Thr]). BICD2 is a golgin and motor-adaptor protein involved in Golgi dynamics and vesicular and mRNA transport. Transient transfection of HeLa cells with all three mutant BICD2 cDNAs caused massive Golgi fragmentation. This observation was even more prominent in primary fibroblasts from an individual harboring c.2108C>T (p.Thr703Met) (affecting the C-terminal coiled-coil domain) and slightly less evident in individuals with c.563A>C (p.Asn188Thr) (affecting the N-terminal coiled-coil domain). Furthermore, BICD2 levels were reduced in affected individuals and trapped within the fragmented Golgi. Previous studies have shown that Drosophila mutant BicD causes reduced larvae locomotion by impaired clathrin-mediated synaptic endocytosis in neuromuscular junctions. These data emphasize the relevance of BICD2 in synaptic-vesicle recycling and support the conclusion that BICD2 mutations cause congenital slowly progressive dominant SMA.  相似文献   
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竞争和非生物胁迫影响处于地理分布边界的红树植物的个体大小 关于红树植物竞争的研究大多局限于幼苗和人工林。我们首次对天然红树林中成年红树的种内竞争进行了控制实验研究,旨在检验竞争和非生物因子在决定红树植物个体大小中的相对重要性。研究样 地位于靠近红树林地理分布边界的美国德克萨斯州阿兰萨斯港(Port Aransas)附近区域。该区域的红树林由“灌丛”状的黑红树(萌芽白骨壤,Avicennia germinans)单一物种组成。我们对10个样方中原生红树 林进行疏伐,形成系列红树林覆盖度梯度,在2013–2019年期间观测各样方中红树植物的生长指标,量化分析红树林覆盖度对红树植物生长的影响;并于2019年调查了红树林的冠层高度。研究结果表明,在该研究期间,红树植物的相对生长速率随着红树林覆盖度的增加而降低,100%红树林覆盖度样方中的红 树植物大小几乎没有增长,说明它们已经达到了该红树林密度条件下的最大尺寸。在红树林覆盖度降低 的样方中,株高明显增加,在红树林覆盖度为11%的样方中,红树植物株高增加了约52%。对比临水岸 边和林内两种生境中的样方,处于临水岸边生境的红树林冠层高度比处于林内生境的高约30%,且这两 种生境的红树林冠层高度均随红树林覆盖度的增加而降低。叶片叶绿素含量和冠层光截留量的测定数据 显示,该区域红树植物的生长也受到氮限制的影响。由此表明,处于地理分布边界的“灌丛”状红树林一 方面受到营养的限制,另一方面红树植物种内个体间仍存在较为强烈的竞争,且种内竞争对红树植物生长的影响较该红树林内非生物生境因子更为重要。  相似文献   
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Understanding gene regulatory networks in mammalian cells requires detailed knowledge of protein-DNA interactions. Commonly used methods for genome-wide mapping of these interactions are based on chromatin immunoprecipitation. However, these methods have some drawbacks, such as the use of crosslinking reagents, the need for highly specific antibodies and relatively large amounts of starting material. We present DamID, an alternative technique to map genome-wide occupancy of interaction sites in vivo, that bypasses these limitations. DamID is based on the expression of a fusion protein consisting of a protein of interest and DNA adenine methyltransferase (Dam). This leads to methylation of adenines near sites where the protein of interest interacts with the DNA. These methylated sequences are subsequently amplified by a methylation-specific PCR protocol and identified by hybridization to microarrays. Using DamID, genome-wide maps of the binding of DNA-interacting proteins in mammalian cells can be constructed efficiently. Depending on the strategy used for expression of the Dam-fusion proteins, genome-wide binding maps can be obtained in as little as 2 weeks.  相似文献   
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Oostdijk  Maartje  Sturlud&#;ttir  Erla  Santos  Maria J. 《Ecosystems》2022,25(5):1117-1134
Ecosystems - The Arctic may be particularly vulnerable to the consequences of both ocean acidification (OA) and global warming, given the faster pace of these processes in comparison with global...  相似文献   
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The advance of metagenomics in combination with intricate cultivation approaches has facilitated the discovery of novel ammonia-, methane-, and other short-chain alkane-oxidizing microorganisms, indicating that our understanding of the microbial biodiversity within the biogeochemical nitrogen and carbon cycles still is incomplete. The in situ detection and phylogenetic identification of novel ammonia- and alkane-oxidizing bacteria remain challenging due to their naturally low abundances and difficulties in obtaining new isolates from complex samples. Here, we describe an activity-based protein profiling protocol allowing cultivation-independent unveiling of ammonia- and alkane-oxidizing bacteria. In this protocol, 1,7-octadiyne is used as a bifunctional enzyme probe that, in combination with a highly specific alkyne-azide cycloaddition reaction, enables the fluorescent or biotin labeling of cells harboring active ammonia and alkane monooxygenases. Biotinylation of these enzymes in combination with immunogold labeling revealed the subcellular localization of the tagged proteins, which corroborated expected enzyme targets in model strains. In addition, fluorescent labeling of cells harboring active ammonia or alkane monooxygenases provided a direct link of these functional lifestyles to phylogenetic identification when combined with fluorescence in situ hybridization. Furthermore, we show that this activity-based labeling protocol can be successfully coupled with fluorescence-activated cell sorting for the enrichment of nitrifiers and alkane-oxidizing bacteria from complex environmental samples, enabling the recovery of high-quality metagenome-assembled genomes. In conclusion, this study demonstrates a novel, functional tagging technique for the reliable detection, identification, and enrichment of ammonia- and alkane-oxidizing bacteria present in complex microbial communities.Subject terms: Environmental microbiology, Sequencing, Microbiology  相似文献   
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