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951.
Michelle L Cloutier A Toutant J Shkreta L Thibault P Durand M Garneau D Gendron D Lapointe E Couture S Le Hir H Klinck R Elela SA Prinos P Chabot B 《Molecular and cellular biology》2012,32(5):954-967
Several apoptotic regulators, including Bcl-x, are alternatively spliced to produce isoforms with opposite functions. We have used an RNA interference strategy to map the regulatory landscape controlling the expression of the Bcl-x splice variants in human cells. Depleting proteins known as core (Y14 and eIF4A3) or auxiliary (RNPS1, Acinus, and SAP18) components of the exon junction complex (EJC) improved the production of the proapoptotic Bcl-x(S) splice variant. This effect was not seen when we depleted EJC proteins that typically participate in mRNA export (UAP56, Aly/Ref, and TAP) or that associate with the EJC to enforce nonsense-mediated RNA decay (MNL51, Upf1, Upf2, and Upf3b). Core and auxiliary EJC components modulated Bcl-x splicing through different cis-acting elements, further suggesting that this activity is distinct from the established EJC function. In support of a direct role in splicing control, recombinant eIF4A3, Y14, and Magoh proteins associated preferentially with the endogenous Bcl-x pre-mRNA, interacted with a model Bcl-x pre-mRNA in early splicing complexes, and specifically shifted Bcl-x alternative splicing in nuclear extracts. Finally, the depletion of Y14, eIF4A3, RNPS1, SAP18, and Acinus also encouraged the production of other proapoptotic splice variants, suggesting that EJC-associated components are important regulators of apoptosis acting at the alternative splicing level. 相似文献
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954.
The lower Mokelumne River (LMR), located in the California Central Valley, supports a population of natural-origin Oncorhynchus mykiss. In addition, the Mokelumne River Fish Hatchery (Hatchery) contributes hatchery produced O. mykiss to the system annually. We conducted a 3 year acoustic tagging study to evaluate the migratory characteristics of LMR hatchery
and natural-origin O. mykiss to the Pacific Ocean. Specifically, we analyzed downstream movement and migration rates, routes, and success of acoustically
tagged O. mykiss of hatchery and natural origin under variable release locations in non-tidal and tidal habitats. Results from our study suggest
there are significant differences in the proportion of hatchery and natural O. mykiss that demonstrate downstream movement. Fish origin, size, and release location all had a significant effect on whether an
individual demonstrated downstream movement. Mokelumne origin O. mykiss that initiated downstream movement utilized numerous migration routes throughout the Delta during their migration towards
the Pacific Ocean. We identified four primary migration pathways from the lower Mokelumne River through the Sacramento-San
Joaquin Delta while the Delta Cross Channel was closed. However, several other pathways were utilized. Origin had a significant
effect on O. mykiss success in reaching key points in the Delta and through the Estuary. Fish size had a significant effect on whether an individual
reached the marine environment. Of the 467 O. mykiss tagged, 34 successfully reached the Pacific Ocean (Golden Gate Bridge), and of these, 33 were hatchery-origin and 1 was natural-origin.
A higher proportion of hatchery-origin fish (10% of tagged) migrated to the ocean compared to natural-origin fish (<1%). Our
study provides valuable information on the differences between hatchery and natural-origin O. mykiss migration characteristics as well as unique insight into the migratory behavior of little studied non-Sacramento River origin
salmonids. 相似文献
955.
NJ Krautler V Kana J Kranich Y Tian D Perera D Lemm P Schwarz A Armulik JL Browning M Tallquist T Buch JB Oliveira-Martins C Zhu M Hermann U Wagner R Brink M Heikenwalder A Aguzzi 《Cell》2012,150(1):194-206
The differentiation of follicular dendritic cells (FDC) is essential to the remarkable microanatomic plasticity of lymphoid follicles. Here we show that FDC arise from ubiquitous perivascular precursors (preFDC) expressing platelet-derived growth factor receptor β (PDGFRβ). PDGFRβ-Cre-driven reporter gene recombination resulted in FDC labeling, whereas conditional ablation of PDGFRβ(+)-derived cells abolished FDC, indicating that FDC originate from PDGFRβ(+) cells. Lymphotoxin-α-overexpressing prion protein (PrP)(+) kidneys developed PrP(+) FDC after transplantation into PrP(-) mice, confirming that preFDC exist outside lymphoid organs. Adipose tissue-derived PDGFRβ(+) stromal-vascular cells responded to FDC maturation factors and, when transplanted into lymphotoxin β receptor (LTβR)(-) kidney capsules, differentiated into Mfge8(+)CD21/35(+)FcγRIIβ(+)PrP(+) FDC capable of trapping immune complexes and recruiting B cells. Spleens of lymphocyte-deficient mice contained perivascular PDGFRβ(+) FDC precursors whose expansion required both lymphoid tissue inducer (LTi) cells and lymphotoxin. The ubiquity of preFDC and their strategic location at blood vessels may explain the de novo generation of organized lymphoid tissue at sites of lymphocytic inflammation. 相似文献
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Truncation of the algal light-harvesting antenna is expected to enhance photosynthetic productivity. The wild type and three mutant strains of Synechocystis sp. strain 6803 with a progressively smaller phycobilisome antenna were examined under different light and CO(2) conditions. Surprisingly, such antenna truncation resulted in decreased whole-culture productivity for this cyanobacterium. 相似文献
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959.
Fidopiastis PM Rader BA Gerling DG Gutierrez NA Watkins KH Frey MW Nyholm SV Whistler CA 《Journal of bacteriology》2012,194(15):3995-4002
Vibrio fischeri cells are the sole colonists of a specialized light organ in the mantle cavity of the sepiolid squid Euprymna scolopes. The process begins when the bacteria aggregate in mucus secretions outside the light organ. The cells eventually leave the aggregate, enter the light organ, and encounter a rich supply of peptides. The need to dissociate from mucus and presumably utilize peptides led us to hypothesize that protease activity is integral to the colonization process. Protease activity associated with whole cells of Vibrio fischeri strain ES114 was identified as the product of a putative cell membrane-associated aminopeptidase (PepN). To characterize this activity, the aminopeptidase was cloned, overexpressed, and purified. Initial steady-state kinetic studies revealed that the aminopeptidase has broad activity, with a preference for basic and hydrophobic side chains and k(cat) and K(m) values that are lower and smaller, respectively, than those of Escherichia coli PepN. A V. fischeri mutant unable to produce PepN is significantly delayed in its ability to colonize squid within the first 12 h, but eventually it establishes a wild-type colonization level. Likewise, in competition with the wild type for colonization, the mutant is outcompeted at 12 h postinoculation but then competes evenly by 24 h. Also, the PepN-deficient strain fails to achieve wild-type levels of cells in aggregates, suggesting an explanation for the initial colonization delay. This study provides a foundation for more studies on PepN expression, localization, and role in the early stages of squid colonization. 相似文献
960.
Bioluminescence energy transfer (BRET) is a powerful tool for the study of protein-protein interactions and conformational changes within proteins. We directly compared two recently developed variants of Renilla luciferase (RLuc), RLuc2 and RLuc8, as BRET donors using an in vitro thrombin assay. The comparison was carried out by placing a thrombin-specific cleavage sequence between the donor luciferase and a green fluorescent protein (GFP(2)) acceptor. Substitution of native RLuc with the RLuc mutants, RLuc2 and 8, in a BRET(2) fusion protein increased the light output by a factor of ~10. Substitution of native RLuc with either of the RLuc mutants resulted in a decrease in BRET(2) ratio by a factor of ~2 when BRET(2) components were separated by the thrombin cleavage sequence. BRET(2) ratios changed by factors of 18.8±1.2 and 18.2±0.4 for GFP(2)-RG-RLuc2 and GFP(2)-RG-RLuc8 fusion proteins, respectively, on thrombin cleavage compared to 28.8±0.20 for GFP(2)-RG-RLuc. The detection limits for thrombin were 0.23 and 0.26 nM for RLuc2 and RLuc8 BRET(2) systems, respectively, and 15 pM for GFP(2)-RG-RLuc. However, overall, the mutant BRET systems remain more sensitive than FRET and brighter than standard BRET(2). 相似文献