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941.
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
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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
942.
Crystallization of the first three domains of the human insulin-like growth factor-1 receptor.
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N. M. McKern M. Lou M. J. Frenkel A. Verkuylen J. D. Bentley G. O. Lovrecz N. Ivancic T. C. Elleman T. P. Garrett L. J. Cosgrove C. W. Ward 《Protein science : a publication of the Protein Society》1997,6(12):2663-2666
The insulin-like growth factor-1 receptor (IGF-1R) is a tyrosine kinase receptor of central importance in cell proliferation. A fragment (residues 1-462) comprising the L1-cysteine rich-L2 domains of the human IGF-1R ectodomain has been overexpressed in glycosylation-deficient Lec8 cells and has been affinity-purified via a c-myc tag followed by gel filtration. The fragment was recognized by two anti-IGF-1R monoclonal antibodies, 24-31 and 24-60, but showed no detectable binding of IGF-1 or IGF-2. Isocratic elution of IGF-1R/462 on anion-exchange chromatography reduced sample heterogeneity, permitting the production of crystals that diffracted to 2.6 A resolution with cell dimensions a = 77.0 A, b = 99.5 A, c = 120.1 A, and space group P2(1)2(1)2(1). 相似文献
943.
Design of a leucine zipper coiled coil stabilized 1.4 kcal mol-1 by phosphorylation of a serine in the e position.
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L. Szilk J. Moitra C. Vinson 《Protein science : a publication of the Protein Society》1997,6(6):1273-1283
Using a dimeric bZIP protein, we have designed a leucine zipper that becomes more stable after a serine in the e position is phosphorylated by protein kinase A (delta delta GP = -1.4 kcal mol-1 dimer-1 or -0.7 kcal mol-1 residue-1). Mutagenesis studies indicate that three arginines form a network of inter-helical (i,i' + 5; i, i' + 2) and intra-helical (i, i + 4) attractive interactions with the phosphorylated serine. When the arginines are replaced with lysines, the stabilizing effect of serine phosphorylation is reduced (delta delta GP = -0.5 kcal mol-1 dimer-1). The hydrophobic interface of the leucine zipper needs a glycine in the d position to obtain an increase in stability after phosphorylation. The phosphorylated protein binds DNA with a 15-fold higher affinity. Using a transient transfection assay, we document a PKA dependent four-fold activation of a reporter gene. Phosphorylation of a threonine in the same e position decreases the stability by delta delta GP = +1.2 kcal mol-1 dimer-1. We present circular dichroism (CD) thermal denaturations of 15 bZIP proteins before and after phosphorylation. These data provide insights into the structural determinants that result in stabilization of a coiled coil by phosphorylation. 相似文献
944.
Akira Uchimura Toshiyuki Shimizu Masahiro Morita Hitomi Ueno Kazuhiro Motoki Hideaki Fukushima Takenori Natori Yasuhiko Koezuka 《Bioorganic & medicinal chemistry》1997,5(12):2245-2249
We compared the immunostimulatory effects of chemically synthesized α-galactosylceramides (α-GalCers), α-glucosylceramides (α-GluCers), 6″-monoglycosylated α-GalCer and 6″- or 4″-monoglycosylated α-GluCer and made the following observations: (1) the length of the fatty acid side chain in the ceramide portions greatly affects the immunostimulatory effects of α-GalCers and α-GluCers; (2) the configuration of the 4″-hydroxyl group of the inner pyranose moiety plays an important role in the immunostimulatory effects of monoglycosylated α-
-pyranosylceramides; (3) the free 4″-hydroxyl group of the inner pyranose of monoglycosylated α-
-pyranosylceramides plays a more important role in their immunostimulatory effects than the free 6″-hydroxyl group. 相似文献
945.
对我国文献中记载的前原鹅观草(Roegneriamayebarana(Honda)Ohwi)的标本和植物,与原产于日本的该种进行了比较形态学、细胞学研究,二者差异显著。作者认为我国所记载的该种种名应是山东鹅观草(Roegneriashandongensis(B.Salomon)J.L.Yang,Y.H.ZhouetYen),其内稃先端钝圆,长为外稃的3/4,染色体数为2n=4x=28,具SY染色体组,结实率达90%以上,过去被错定为R.mayebarana。而R.mayebarana在日本系一天然杂种,其内稃先端尖,与外稃等长或稍短,染色体数为2n=6x=42,具HSY染色体组,结实率极低,仅0.2%~0.4%。 相似文献
946.
947.
EB病毒BNLF-1基因的分子生物学研究进展 总被引:2,自引:0,他引:2
位于EB病毒基因组U5-TR区内的BNLF-1基因,其转译产物为潜伏膜蛋白(latent membrane protein1, LMP-1),由于LMP-1可以导致细胞转化并在EB病毒致癌过程中具有重要作用,因而成为近年来EB病毒分子生物学及相关肿瘤如人鼻咽癌、伯基特淋巴瘤、何杰金氏病等疾病病因发病学研究的热点,并取得了一批有重要意义的成果,文章从BNLF-1的基因结构及表达调控, LMP蛋白的结构及生化功能, LMP-1的生物学功能和LMP-1研究进行评述. 相似文献
948.
Kenji Hara Henneke Pangkey Kiyoshi Osatomi Keiko Yatsuda Atsushi Hagiwara Katsuyasu Tachibana Tadashi Ishihara 《Hydrobiologia》1997,358(1-3):89-94
We examined some characteristics of hydrolyticenzymes, especially -1,3-glucanase, to obtain theinformation of cell wall lytic enzymes forrotifers.Crude enzyme (ammonium sulfate fraction) of rotifershydrolyzed starch, -1,3-glucan, glycol chitinand CM-cellulose. Optimum pH for hydrolysis ofstarch and CM-cellulose was 6.5, and that for -1,3glucan and glycol chitin was pH 6.0. Pectic acid,xylan and agarose were not hydrolyzed at pH 3–10.-1,3 glucanase was purified about 73-fold from crudeenzyme by ion-exchange chromatography and gelfiltration. Optimum pH and temperature of the enzymewere 6 and 60 °C, respectively. The molecular weight ofthe enzyme was estimated about 260 kDa by gelfiltration. The enzyme was inhibited byHgCl2 and MnCl_2. 相似文献
949.
H.C. Chan S.H. Law P.S. Leung L.X.M. Fu P.Y.D. Wong 《The Journal of membrane biology》1997,156(3):241-249
The β-adrenergic (cAMP-dependent) regulation of Cl− conductance is defective in cystic fibrosis (CF). The present study explored alternative regulation of anion secretion in
CF pancreatic ductal cells (CFPAC-1) by angiotensin II (AII) using the short-circuit current (I
SC
) technique. An increase in I
SC
could be induced in CFPAC-1 cells by basolateral or apical application of AII in a concentration-dependent manner (EC50 at 3 μm and 100 nm, respectively). Angiotensin receptor subtypes were identified using specific antagonists, losartan and PD123177, for AT1 and AT2 receptors, respectively. It was found that losartan (1 μm) could completely inhibit the AII-induced I
SC
, whereas, PD123177 exerted insignificant effect on the I
SC
, indicating predominant involvement of AT1 receptors. The presence of AT1 receptors in CFPAC-1 cells was also demonstrated by immunohistochemical studies using specific antibodies against AT1 receptors. Confocal microscopic study demonstrated a rise in intracellular Ca2+ upon stimulation by AII indicating a role of intracellular Ca2+ in mediating the AII response. Depletion of intracellular but not extracellular pool of Ca2+ diminished the AII-induced I
SC
. Treatment of the monolayers with a Cl− channel blocker, DIDS, markedly reduced the I
SC
, indicating that a large portion of the AII-activated I
SC
was Cl−-dependent. AII-induced I
SC
was also observed in monolayers whose basolateral membranes had been permeabilized by nystatin, suggesting that the I
SC
was mediated by apical Cl− channels. Our study indicates an AT1-mediated Ca2+-dependent regulatory mechanism for anion secretion in CF pancreatic duct cells which may be important for the physiology
and pathophysiology of the pancreas.
Received: 17 June 1996/Revised: 14 November 1996 相似文献
950.
We here report on studies on the frog skin epithelium to identify the nature of its excretory H+ pump by comparing transport studies, using inhibitors highly specific for V-ATPases, with results from immunocytochemistry
using V-ATPase-directed antibodies. Bafilomycin A1 (10 μm) blocked H+ excretion (69 ± 8% inhibition) and therefore Na+ absorption (61 ± 17% inhibition after 60 min application, n= 6) in open-circuited skins bathed on their apical side with a 1 mm Na2SO4 solution, ``low-Na+ conditions' under which H+ and Na+ fluxes are coupled 1:1. The electrogenic outward H+ current measured in absence of Na+ transport (in the presence of 50 μm amiloride) was also blocked by 10 μm bafilomycin A1 or 5 μm concanamycin A. In contrast, no effects were found on the large and dominant Na+ transport (short-circuit current), which develops with apical solutions containing 115 mm Na+ (``high-Na+ conditions'), demonstrating a specific action on H+ transport. In immunocytochemistry, V-ATPase-like immunoreactivity to the monoclonal antibody E11 directed to the 31-kDa subunit
E of the bovine renal V-ATPase was localized only in mitochondria-rich cells (i) in their apical region which corresponds
to apical plasma membrane infoldings, and (ii) intracellularly in their neck region and apically around the nucleus. In membrane
extracts of the isolated frog skin epithelium, the selectivity of the antibody binding was tested with immunoblots. The antibody
labeled exclusively a band of about 31 kDa, very likely the corresponding subunit E of the frog V-ATPase. Our investigations
now deliver conclusive evidence that H+ excretion is mediated by a V-ATPase being the electrogenic H+ pump in frog skin.
Received: 21 May 1996/Revised: 24 December 1996 相似文献