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991.
Melissa A. Kofoed David A. Wampler Arti S. Pandey John W. Peters Scott A. Ensign 《Journal of bacteriology》2011,193(18):4904-4913
NADPH:2-ketopropyl-coenzyme M oxidoreductase/carboxylase (2-KPCC), an atypical member of the disulfide oxidoreductase (DSOR) family of enzymes, catalyzes the reductive cleavage and carboxylation of 2-ketopropyl-coenzyme M [2-(2-ketopropylthio)ethanesulfonate; 2-KPC] to form acetoacetate and coenzyme M (CoM) in the bacterial pathway of propylene metabolism. Structural studies of 2-KPCC from Xanthobacter autotrophicus strain Py2 have revealed a distinctive active-site architecture that includes a putative catalytic triad consisting of two histidine residues that are hydrogen bonded to an ordered water molecule proposed to stabilize enolacetone formed from dithiol-mediated 2-KPC thioether bond cleavage. Site-directed mutants of 2-KPCC were constructed to test the tenets of the mechanism proposed from studies of the native enzyme. Mutagenesis of the interchange thiol of 2-KPCC (C82A) abolished all redox-dependent reactions of 2-KPCC (2-KPC carboxylation or protonation). The air-oxidized C82A mutant, as well as wild-type 2-KPCC, exhibited the characteristic charge transfer absorbance seen in site-directed variants of other DSOR enzymes but with a pKa value for C87 (8.8) four units higher (i.e., four orders of magnitude less acidic) than that for the flavin thiol of canonical DSOR enzymes. The same higher pKa value was observed in native 2-KPCC when the interchange thiol was alkylated by the CoM analog 2-bromoethanesulfonate. Mutagenesis of the flavin thiol (C87A) also resulted in an inactive enzyme for steady-state redox-dependent reactions, but this variant catalyzed a single-turnover reaction producing a 0.8:1 ratio of product to enzyme. Mutagenesis of the histidine proximal to the ordered water (H137A) led to nearly complete loss of redox-dependent 2-KPCC reactions, while mutagenesis of the distal histidine (H84A) reduced these activities by 58 to 76%. A redox-independent reaction of 2-KPCC (acetoacetate decarboxylation) was not decreased for any of the aforementioned site-directed mutants. We interpreted and rationalized these results in terms of a mechanism of catalysis for 2-KPCC employing a unique hydrophobic active-site architecture promoting thioether bond cleavage and enolacetone formation not seen for other DSOR enzymes. 相似文献
992.
Benjamin Wipfler Ward Koehler Paul B. Frandsen Alexander Donath Shanlin Liu Ryuichiro Machida Bernhard Misof Ralph S. Peters Shota Shimizu Xin Zhou Sabrina Simon 《Systematic Entomology》2020,45(3):516-526
Earwigs are one of the comparatively species-poor insect orders. Although various aspects of the phylogeny of this lineage are poorly understood, before the present study, there was a general consensus that Dermaptera comprises two major lineages: the paraphyletic Protodermaptera or ‘lower earwigs’ and the monophyletic Epidermaptera or ‘higher earwigs’, which are nested within the former. Our phylogenomic study based on the analysis of 3247 nuclear single-copy genes reverses these relationships by placing monophyletic Protodermaptera within paraphyletic Epidermaptera. This phylogenetic reversal among the major earwig lineages is not contradicted by morphological arguments but results in far-reaching reinterpretations of the dermapteran ground plan. Within Dermaptera, Apachyidae form the sister group to the remaining earwigs which might imply that social behaviour is not part of the earwig ground plan. Our results corroborate the monophyly of Eudermaptera within Epidermaptera and the paraphyly of several traditional families. The monophyly of Protodermaptera is supported by molecular and morphological evidence, although the exact position of Karschiellidae which were not included in the molecular dataset cannot be determined. 相似文献
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995.
The following high performance liquid chromatography system was found suitable for separating most lipoxygenase metabolites of arachidonic acid: Techsphere 5-C18 column, eluting solvent methanol:water:acetic acid (65:35:0.06 v/v), pH 5.3. Comparisons with other packing materials and solvent systems are described. The method could be used to identify lipoxygenase products released from mouse macrophage cells stimulated with gamma-hexachlorocyclohexane. Detection limits between 1 and 10 ng were obtained. 相似文献
996.
Anostracans were found living in ephemeral pools in the dark sections of three caves on the As Summan Plateau in Saudi Arabia.
Branchipus schaefferi Fischer, 1834 occurred alone in one while it cohabited with Streptocephalus torvicornisbucheti Daday,
1910 in a second cave; fairy shrimps were observed but not collected from the third. None of the specimens demonstrated any
of the types of morphological changes typically associated with cave adapted species. This is likely due to continuing colonization
of the pools during flooding events.
This revised version was published online in August 2006 with corrections to the Cover Date. 相似文献
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In this study 10 patients with acute myelocytic leukemia (AML) each received a rapid intravenous injection of high dose cytosine arabinoside (HD Ara-C; 1 g/m2). Bone marrow aspirates were obtained before and after Ara-C administration to determine the percentage of cells in S-phase measured by flow cytometry. In 5 out of 10 cases synchronization of the leukemic cells in S-phase of the cell cycle was observed. However, the time of maximum synchronization turned out to be difficult to predict. Therefore, the strong correlation between percentage of cells in S-phase at diagnosis and the time of maximal accumulation of S-phase cells after Ara-C administration, as observed by others in childhood AML, could not be confirmed for adult AML patients. Although synchronization of AML cells after in vivo Ara-C administration could be demonstrated in at least half of the patients, the practical consequences are such that clinical application was hampered. 相似文献
1000.
C Kozak G Peters R Pauley V Morris R Michalides J Dudley M Green M Davisson O Prakash A Vaidya et al. 《Journal of virology》1987,61(5):1651-1654
We propose a revised standardized nomenclature for endogenous mouse mammary tumor viruses based on characterization by molecular cloning techniques and genetic segregation data. 相似文献