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51.
G. Gradl P. Grandison E. Lindridge Y. Wang J. Watson F. Rudert 《Apoptosis : an international journal on programmed cell death》1996,1(2):131-140
Different CD95 (Fas/APO-1) isoforms and phosphory lated CD95 species were identified in human T and B cell lines. We had shown
previously that the CD95 intracellular domain (IC), expressed as a glutathione S-transferase (GST) fusion protein in murine
L929 fibroblasts, was phosphorylatedin vivo. GST-CD95IC was phosphorylatedin vitro by a kinase present in extracts from the human lymphocytic cell lines Jurkat and MP-1 and from murine L929 cells. Phosphoamino
acid analysis indicated that phosphorylation occurred at multiple threonine residues and also at tyrosine (Tyr232 and Tyr291)
and serine. Amino acids 191 to 275 of CD95 were sufficient for phosphorylation at threonine, tyrosine and serine and also
mediated interaction with a 35 kDa cellular protein. Immuno-precipitation of CD95 and chemical cross-linking revealed CD95-associated
proteins of approximately 35, 45 and 75 kDa. GST-CD95IC affinity chromatography detected binding of the 35 and 75 kDa protein
species. The 75 kDa species may correspond to the CD95-associated proteins RIP or FAF1 and the 35 kDa protein may represent
a TRADD analogue. These data indicate that several cellular proteins interact with CD95, possibly in a multi-protein complex,
and that a kinase activity is associated with CD95 not onlyin vitro but alsoin vivo. Therefore, receptor phosphorylation may play a role in CD95 signal transduction.
This work was in part supported by a grant from the Health Research Council of New Zealand (to JW). 相似文献
52.
X Xiao G Hintermann AL Demanin J Piret 《Journal of industrial microbiology & biotechnology》1996,16(4):261-262
Streptomyces glaucescens is shown to possess -lactamase activity which is inhibitable by clavulanate. This is important in regard to its use as a cloning host for enzymes of \-lactam biosynthesis. 相似文献
53.
54.
利用单克隆抗体免疫磁珠吸附方法分离脐血CD34+细胞,并观察了IL3/GMCSF融合蛋白(PIXY321)对脐血CD34+细胞的刺激作用。PIXY321对脐血CD34+细胞扩增作用大于IL3和GMCSF单独及联合应用。在液体培养条件下,每毫升20ngPIXY321可有效地扩增脐血造血祖细胞,适宜扩增时间为5-8天,扩增后造血祖细胞的数量可达扩增前的8-10倍,从而初步建立了一种简单可行的脐血造血细胞扩增方法。 相似文献
55.
本研究用CD23单克隆抗体交联活化的人扁桃体B细胞,证明CD23McAb对B细胞呈双向调节效应,即:高浓度区抑制B细胞增殖,低浓度区促进B细胞增殖,继而,用对B细胞有抑制效应浓度的CD23McAb交联B细胞膜CD23分子,通过研究抑制效应恢复条件,探讨了CD23McAb产生抑制效应的作用机制。结果显示:去除CD23McAb后,抑制作用不能恢复,用SAC再次活化,使B细胞恢复增殖状态,用促B细胞增殖 相似文献
56.
Nishijima K Hisatsune T Kato H Kohyama M Kakehi M Hachimura S Kaminogawa S 《Cytotechnology》1997,25(1-3):89-100
Feeding of a whole casein diet, which abolished the αs1-casein-specific proliferation and IFN-γ productivity of CD4+ T cells, did not affect the proliferative response of CD8+ T cells with regard to the antigen dose response, cell dose response, kinetics of the proliferation and epitope specificity,
as well as IFN-γ production. To assess the characteristics of the CD8+ T cells, we established αs1-casein-specific CD8+ T cell clones from both casein-fed and control mice. The established clones produced different amount of IFN-γ and IL-10,
and one clone derived from the casein-fed mice produced a remarkable amount of IL-10. The clones from casein-fed mice produced
considerable amounts of TGF-β, while those from control mice produced only small amounts. The possible role of CD8+ T cells in oral tolerance is discussed.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
57.
François-Yves Dupradeau Guillaume Le Flem Jean-Michel Wieruszeski Manuel Dauchez Alain Alix Véronique Larreta-Garde Jean-Pierre Monti 《Letters in Peptide Science》1997,4(4-6):489-495
H-NMR studies of the bovine insulin S-sulfonatedB-chain are reported in H2O/D2O (9/1) and inglycerol-d5 (5 M) using two-dimensional NMRspectroscopy. The first results show that the oxidizedinsulin B-chain secondary structure differs from thatof native insulin by a loss of the -helixbetween the two disulfide bridges and that theglycerol favours the structuring of the peptide. 相似文献
58.
The role of context on alpha-helix stabilization: host-guest analysis in a mixed background peptide model. 总被引:1,自引:1,他引:0 下载免费PDF全文
J. Yang E. J. Spek Y. Gong H. Zhou N. R. Kallenbach 《Protein science : a publication of the Protein Society》1997,6(6):1264-1272
The helix content of a series of peptides containing single substitutions of the 20 natural amino acids in a new designed host sequence, succinyl-YSEEEEKAKKAXAEEAEKKKK-NH2, has been determined using CD spectroscopy. This host is related to one previously studied, in which triple amino acid substitutions were introduced into a background of Glu-Lys blocks completely lacking alanine. The resulting free energies show that only Ala and Glu- prove to be helix stabilizing, while all other side chains are neutral or destabilizing. This agrees with results from studies of alanine-rich peptide modela, but not the previous Glu-Lys block oligomers in which Leu and Met also stabilize helix. The helix propensity scale derived from the previous block oligomers correlated well with the frequencies of occurrence of different side chains in helical sequences of proteins, whereas the values from the present series do not. The role of context in determining scales of helix propensity values is discussed, and the ability of algorithms designed to predict helix structure from sequence is compared. 相似文献
59.
Design, synthesis, expression, and characterization of the genes for mouse Fc gamma RIIb1 and Fc gamma RIIb2 cytoplasmic regions. 下载免费PDF全文
L. Chen N. L. Thompson G. J. Pielak 《Protein science : a publication of the Protein Society》1997,6(5):1038-1046
The cytoplasmic regions of the mouse low-affinity Fc gamma RII isoforms, mFc gamma RIIb1, and mFc gamma RIIb2, play a key role in signal transduction by mediating different cellular functions. mFc gamma RIIb1 has a 94-residue cytoplasmic region, whereas mFc gamma RIIb2 has a 47-residue cytoplasmic region. Genes encoding the cytoplasmic regions of mFc gamma RIIb1 (b1-94) and mFc gamma RIIb2 (b2-47) were designed, synthesized, and expressed as fusion proteins in Escherichia coli. A sequence-specific protease, thrombin, was used to release the b1-94 peptide, which was purified by using HPLC. The b2-47 peptide was synthesized chemically. CD spectropolarimetry was employed to examine the secondary structures of b1-94 and b2-47. These studies were conducted in aqueous solution, in mixtures of water and trifluoroethanol or methanol, and as a function of temperature. The results indicate that the b1-94 and b2-47 structures are sensitive functions of the solvent environment, and that nonaqueous solvents induce significant alpha-helical structure. 相似文献
60.
Probing minimal independent folding units in dihydrofolate reductase by molecular dissection. 总被引:7,自引:5,他引:2 下载免费PDF全文
C. V. Gegg K. E. Bowers C. R. Matthews 《Protein science : a publication of the Protein Society》1997,6(9):1885-1892
Molecular dissection was employed to identify minimal independent folding units in dihydrofolate reductase (DHFR) from Escherichia coli. Eight overlapping fragments of DHFR, spanning the entire sequence and ranging in size from 36 to 123 amino acids, were constructed by chemical cleavage. These fragments were designed to examine the effect of tethering multiple elements of secondary structure on folding and to test if the secondary structural domains represent autonomous folding units. CD and fluorescence spectroscopy demonstrated that six fragments containing up to a total of seven alpha-helices or beta-strands and, in three cases, the adenine binding domain (residues 37-86), are largely disordered. A stoichiometric mixture of the two fragments comprising the large discontinuous domain, 1-36 and 87-159, also showed no evidence for folding beyond that observed for the isolated fragments. A fragment containing residues 1-107 appears to have secondary and tertiary structure; however, spontaneous self-association made it impossible to determine if this structure solely reflects the behavior of the monomeric form. In contrast, a monomeric fragment spanning residues 37-159 possesses significant secondary and tertiary structure. The urea-induced unfolding of fragment 37-159 in the presence of 0.5 M ammonium sulfate was found to be a well-defined, two-state process. The observation that fragment 37-159 can adopt a stable native fold with unique, aromatic side-chain packing is quite striking because residues 1-36 form an integral part of the structural core of the full-length protein. 相似文献