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101.
Wawrzynski A Ashfield T Chen NW Mammadov J Nguyen A Podicheti R Cannon SB Thareau V Ameline-Torregrosa C Cannon E Chacko B Couloux A Dalwani A Denny R Deshpande S Egan AN Glover N Howell S Ilut D Lai H Del Campo SM Metcalf M O'Bleness M Pfeil BE Ratnaparkhe MB Samain S Sanders I Ségurens B Sévignac M Sherman-Broyles S Tucker DM Yi J Doyle JJ Geffroy V Roe BA Maroof MA Young ND Innes RW 《Plant physiology》2008,148(4):1760-1771
Retrotransposons and their remnants often constitute more than 50% of higher plant genomes. Although extensively studied in monocot crops such as maize (Zea mays) and rice (Oryza sativa), the impact of retrotransposons on dicot crop genomes is not well documented. Here, we present an analysis of retrotransposons in soybean (Glycine max). Analysis of approximately 3.7 megabases (Mb) of genomic sequence, including 0.87 Mb of pericentromeric sequence, uncovered 45 intact long terminal repeat (LTR)-retrotransposons. The ratio of intact elements to solo LTRs was 8:1, one of the highest reported to date in plants, suggesting that removal of retrotransposons by homologous recombination between LTRs is occurring more slowly in soybean than in previously characterized plant species. Analysis of paired LTR sequences uncovered a low frequency of deletions relative to base substitutions, indicating that removal of retrotransposon sequences by illegitimate recombination is also operating more slowly. Significantly, we identified three subfamilies of nonautonomous elements that have replicated in the recent past, suggesting that retrotransposition can be catalyzed in trans by autonomous elements elsewhere in the genome. Analysis of 1.6 Mb of sequence from Glycine tomentella, a wild perennial relative of soybean, uncovered 23 intact retroelements, two of which had accumulated no mutations in their LTRs, indicating very recent insertion. A similar pattern was found in 0.94 Mb of sequence from Phaseolus vulgaris (common bean). Thus, autonomous and nonautonomous retrotransposons appear to be both abundant and active in Glycine and Phaseolus. The impact of nonautonomous retrotransposon replication on genome size appears to be much greater than previously appreciated. 相似文献
102.
David M. Burns Yun-Long Li Eric Shi Chunhong He Meizhong Xu Jincong Zhuo Colin Zhang Ding-Quan Qian Yanlong Li Richard Wynn Maryanne B. Covington Kamna Katiyar Cindy A. Marando Jordan S. Fridman Peggy Scherle Steve Friedman Brian Metcalf Wenqing Yao 《Bioorganic & medicinal chemistry letters》2009,19(13):3525-3530
A serendipitous discovery that the metalloprotease binding profile of a novel class of 2-carboxamide-3-hydroxamic acid piperidines could be significantly attenuated by the modification of the unexplored P1 substituent enabled the design and synthesis of a novel 2-carboxamide-1-hydroxamic acid cyclohexyl scaffold core that exhibited excellent HER-2 potency and unprecedented MMP-selectivity that we believe would not have been possible via conventional P1′ perturbations. 相似文献
103.
104.
目的: 在完成吸入室内空气状态下症状限制性最大极限心肺运动试验(CPET)和动脉血气指标动态变化规律的基础上,进一步探讨体液酸碱度和CO2含量对呼吸调控的影响。方法: 选正常志愿者5名,给予5%NaHCO3(总量0.3 g/kg)分次口服,每5 min口服75 ml(3.75g )。总量服完1 h后,重复CPET。于静息、热身、运动及恢复期,连续测定肺通气指标及每分钟动脉取样的血气指标变化,并与本人在非碱化血液条件下对照数据进行配对t检验比较。结果: 碱化血液之后,CPET期间随着运动功率逐步递增,气体交换和血气指标的反应模式与非碱化血液对照相似(P>0.05);即与静息状态比较,每分通气量、潮气量、呼吸频率、VO2、VCO2均呈现近于线性渐进性递增(P<0.05~0.001)。与碱化血液前吸入室内空气的对照比较:在碱化血液条件下,所有时间点血红蛋白浓度,PaCO2与pH均显著提高(P<0.05);除无氧阈PaCO2减低外,只有热身状态呈增高态势,统计学有显著差异(P<0.05);而PaO2无差异(P>0.05),各状态均较对照状态减低,除恢复期外均有统计学差异(P<0.05)。与非碱化血液对照比较,除静息每分通气量低于对照(P<0.05)外,所有通气指标均无统计学差异(P>0.05)。结论: 碱化血液条件下, 尽管有更高的CaCO2, PaCO2 和 pHa平均水平及更低的Hba和[H+]a平均水平,机体对CPET的呼吸反应模式基本相似。 相似文献
105.
目的: 在急性血液碱化前、后空气吸入下完成症状限制性最大极限心肺运动试验(CPET)的基础上,本文探讨在血液碱化后吸入纯氧对呼吸调控的影响。方法: 正常志愿者5名在碱化血液后呼吸纯氧CPET,在静息、热身、运动及恢复期,连续测定肺通换气指标及每分钟动脉取样的血气指标,对CPET期间的呼吸气体交换和血气指标的动态变化进行分析,同时与急性碱化血液前、后空气CPET数据比较。结果: 碱化血液后吸入纯氧运动呼吸反应与急性碱化血液前、后空气CPET呼吸反应基本一致。CPET期间,各运动状态下的每分通气量均与对照组相似(P>0.05);仅静息每分通气量较血液碱化空气CPET略高(P<0.05),而其它状态和恢复2min时均相近(P>0.05)。潮气量仅峰值运动时较对照和血液碱化空气CPET略低(P<0.05);而运动过程和恢复2min时的潮气量均相近(P>0.05)。呼吸频率在各个时间与血液碱化前后CPET均无差异(P>0.05)。在碱化血液后吸入纯氧运动各个时期的PaO2和SaO2较碱化血液前后空气CPET时明显提高(P<0.001,P<0.05)。血红蛋白浓度虽然较急性血液碱化前后均低,但仅较血液碱化前显著降低(P<0.05),比血液碱化后差异不显著(P>0.05) ; 开始时的PaCO2较碱化血液前后空气CPET时降低(P<0.05),无氧阈时相近(P>0.05),但到峰值及恢复2 min时明显增高(P<0.05);pH仅较对照增高(P<0.05),但与碱化血液空气试验时无差异;乳酸水平较对照略高,但仅在热身和恢复期有差异(P<0.05)。纯氧提高了两人无氧阈和三人峰值运动的功率和时间。结论: 虽然血液碱化给予纯氧, CPET呼吸反应与碱化血液前、后空气CPET呼吸反应模式相似,表明运动中呼吸反应主要取决于代谢变化,而非动脉血气平均值高低。 相似文献
106.
James A. Rickard Joanne A. O’Donnell Joseph M. Evans Najoua Lalaoui Ashleigh R. Poh TeWhiti Rogers James E. Vince Kate E. Lawlor Robert L. Ninnis Holly Anderton Cathrine Hall Sukhdeep K. Spall Toby J. Phesse Helen E. Abud Louise H. Cengia Jason Corbin Sandra Mifsud Ladina Di Rago Donald Metcalf Matthias Ernst Grant Dewson Andrew W. Roberts Warren S. Alexander James M. Murphy Paul G. Ekert Seth L. Masters David L. Vaux Ben A. Croker Motti Gerlic John Silke 《Cell》2014
107.
Michael L. Burns Thomas M. Malott Kevin J. Metcalf Benjamin J. Hackel Jonah R. Chan Eric V. Shusta 《Applied and environmental microbiology》2014,80(18):5732-5742
Brain-derived neurotrophic factor (BDNF) plays an important role in nervous system function and has therapeutic potential. Microbial production of BDNF has resulted in a low-fidelity protein product, often in the form of large, insoluble aggregates incapable of binding to cognate TrkB or p75 receptors. In this study, employing Saccharomyces cerevisiae display and secretion systems, it was found that BDNF was poorly expressed and partially inactive on the yeast surface and that BDNF was secreted at low levels in the form of disulfide-bonded aggregates. Thus, for the purpose of increasing the compatibility of yeast as an expression host for BDNF, directed-evolution approaches were employed to improve BDNF folding and expression levels. Yeast surface display was combined with two rounds of directed evolution employing random mutagenesis and shuffling to identify BDNF mutants that had 5-fold improvements in expression, 4-fold increases in specific TrkB binding activity, and restored p75 binding activity, both as displayed proteins and as secreted proteins. Secreted BDNF mutants were found largely in the form of soluble homodimers that could stimulate TrkB phosphorylation in transfected PC12 cells. Site-directed mutagenesis studies indicated that a particularly important mutational class involved the introduction of cysteines proximal to the native cysteines that participate in the BDNF cysteine knot architecture. Taken together, these findings show that yeast is now a viable alternative for both the production and the engineering of BDNF. 相似文献
108.
Peishen Zhao TinaMarie Lieu Nicholas Barlow Matthew Metcalf Nicholas Veldhuis Dane D. Jensen Martina Kocan Silvia Sostegni Silke Haerteis Vera Baraznenok Ian Henderson Erik Lindstr?m Raquel Guerrero-Alba Eduardo E. Valdez-Morales Wolfgang Liedtke Peter McIntyre Stephen J. Vanner Christoph Korbmacher Nigel W. Bunnett 《The Journal of biological chemistry》2014,289(52):35858
109.
Sandra Anne Banack James S. Metcalf Liying Jiang Derek Craighead Leopold L. Ilag Paul Alan Cox 《PloS one》2012,7(11)
Prior to the evolution of DNA-based organisms on earth over 3.5 billion years ago it is hypothesized that RNA was the primary genetic molecule. Before RNA-based organisms arose, peptide nucleic acids may have been used to transmit genetic information by the earliest forms of life on earth. We discovered that cyanobacteria produce N-(2-aminoethyl)glycine (AEG), a backbone for peptide nucleic acids. We detected AEG in axenic strains of cyanobacteria with an average concentration of 1 µg/g. We also detected AEG in environmental samples of cyanobacteria as both a free or weakly bound molecule and a tightly bound form released by acid hydrolysis, at concentrations ranging from not detected to 34 µg/g. The production of AEG by diverse taxa of cyanobacteria suggests that AEG may be a primitive feature which arose early in the evolution of life on earth. 相似文献
110.
Purification of two forms of colony-stimulating factor from mouse L-cell-conditioned medium 总被引:5,自引:0,他引:5
A W Burgess D Metcalf I J Kozka R J Simpson G Vairo J A Hamilton E C Nice 《The Journal of biological chemistry》1985,260(29):16004-16011
A modified procedure for the purification of the colony-stimulating factors (CSFs) in mouse L-cell-conditioned medium is used to isolate two forms of CSF, which are separable by reversed-phase high performance liquid chromatography with 300-A pore size supports. The specific biological activity of these CSFs (2 X 10(9) colonies/mg) was considerably higher than has been achieved by other methods. Even at high concentration (200 pM) both molecules stimulated predominantly more macrophage than granulocyte colonies; however, the less hydrophobic form appeared to stimulate the formation of more pure granulocytic colonies. Almost twice as much of the less hydrophobic CSF was recovered from L-cell-conditioned medium. Analysis using sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated that both forms of L-cell CSF had apparent molecular masses of approximately 70,000 daltons. However, on reduction with 2-mercaptoethanol, while both forms generated a 39,000-dalton subunit, the less hydrophobic form also yielded a 32,000-dalton subunit. Storage of either form of L-cell CSF at pH 2.1, in the presence of acetonitrile or isopropanol, destroyed the biological activity. Electrophoretic analysis of the L-cell CSFs stored under these conditions indicated that this was associated with a spontaneous dissociation of the CSF dimer into the inactive subunits. There was some charge heterogeneity (pI 3.5-4.7) indicating different degrees of glycosylation. The unique N-terminal amino acid sequences of both forms of CSF were the same: (Lys-Glu-Val-Ser-Glu-His-X-Ser-His-Met-Ile-Gly-Asn). Thus, the polypeptide chains appear to be identical for the subunits of both forms of L-cell CSF. 相似文献