全文获取类型
收费全文 | 40219篇 |
免费 | 4196篇 |
国内免费 | 11篇 |
出版年
2023年 | 176篇 |
2022年 | 352篇 |
2021年 | 674篇 |
2020年 | 490篇 |
2019年 | 570篇 |
2018年 | 706篇 |
2017年 | 635篇 |
2016年 | 1051篇 |
2015年 | 1738篇 |
2014年 | 1889篇 |
2013年 | 2344篇 |
2012年 | 2880篇 |
2011年 | 2800篇 |
2010年 | 1860篇 |
2009年 | 1617篇 |
2008年 | 2255篇 |
2007年 | 2246篇 |
2006年 | 2140篇 |
2005年 | 1909篇 |
2004年 | 1905篇 |
2003年 | 1689篇 |
2002年 | 1662篇 |
2001年 | 663篇 |
2000年 | 616篇 |
1999年 | 625篇 |
1998年 | 434篇 |
1997年 | 355篇 |
1996年 | 342篇 |
1995年 | 351篇 |
1994年 | 311篇 |
1993年 | 314篇 |
1992年 | 418篇 |
1991年 | 361篇 |
1990年 | 358篇 |
1989年 | 367篇 |
1988年 | 369篇 |
1987年 | 338篇 |
1986年 | 274篇 |
1985年 | 303篇 |
1984年 | 307篇 |
1983年 | 272篇 |
1982年 | 260篇 |
1981年 | 232篇 |
1980年 | 212篇 |
1979年 | 216篇 |
1978年 | 182篇 |
1977年 | 183篇 |
1976年 | 183篇 |
1975年 | 200篇 |
1973年 | 165篇 |
排序方式: 共有10000条查询结果,搜索用时 15 毫秒
171.
T W Martin D R Feldman K E Goldstein J R Wagner 《Biochemical and biophysical research communications》1989,165(1):319-326
Bovine pulmonary artery endothelial cells (BPAEC) were prelabeled with [3H]choline or [3H]myristic acid to selectively label endogenous phosphatidylcholine. BPAEC were stimulated with ATP and bradykinin (BK), and phospholipase D (PLD) activation was detected as a 4-fold increase in [3H]choline in cells prelabeled with [3H]choline or as a 2- to 3-fold increase in [3H]phosphatidylethanol in cells prelabeled with [3H]myristic acid and stimulated in the presence of ethanol. Pretreatment of BPAEC with 0.1 microM phorbol 12-myristate 13-acetate (PMA) for 22 hr completely inhibited agonist-induced PLD activation, whereas prostacyclin synthesis and [3H]phosphoinositide ([3H]PIns) hydrolysis were enhanced in pretreated cells. Long-term PMA treatment thus dissociates agonist-induced PLD activation from [3H]PIns hydrolysis, and agonist-induced prostacyclin synthesis is not dependent upon PLD activation. 相似文献
172.
Secondary structure and orientation of a chemically synthesized mitochondrial signal sequence in phospholipid bilayers 总被引:2,自引:0,他引:2
E Goormaghtigh I Martin M Vandenbranden R Brasseur J M Ruysschaert 《Biochemical and biophysical research communications》1989,158(2):610-616
A pre-sequence of 25 amino acids is required for import of yeast cytochrome oxidase subunit IV into mitochondria. Structure and orientation of the 25 amino acids synthesized peptide (p25) in a lipid bilayer were investigated by infrared attenuated total reflection spectroscopy. This method allowed to overcome the difficulties related to the optical turbidity due to the light scattering on membrane fragments which prevents the use of circular dichroism. We demonstrate here that incubation of the peptide with DOPC (dioleoylphosphatidylcholine) and DOPC-CL (dioleoylphosphatidylcholine - cardiolipin) liposomes is accompanied by an increase in alpha-helical content as compared to beta structure. Polarisation measurements indicate that the amphipathic helical segment is inserted parallel to the lipid acyl chains in cardiolipin containing liposomes. 相似文献
173.
Martin S?ndergaard 《Hydrobiologia》1989,(1):321
Material cycling
Seasonal variations in the loosely sorbed phosphorus fraction of the sediment of a shallow and hypereutrophic lake 相似文献174.
Walter A. Heidmann Annegret Büthe Martin Beyerbach Reinhard Löhmer und Harald A. Rüssel-Sinn 《Journal of Ornithology》1989,130(3):311-320
Zusammenfassung Rückstände chlorierter Kohlenwasserstoffe in Eiern und Lebern von im Binnenland Niedersachsens brütenden Vogelarten — Feldsperling, Mehlschwalbe, Weißstorch, Graureiher, Saatkrähe, Stockente und andere Arten — werden angegeben und deren Abhängigkeit von Brutort, Nahrung und Zugverhalten diskutiert.
Chlorinated hydrocarbons of some bird species breeding in the inland of Lower Saxony (FRG)
Summary Residues of chlorinated hydrocarbons in eggs and livers of some bird species — Tree Sparrow, House Martin, White Stork, Heron, Rook, Mallard, and further species — are presented. The dependence on place of breeding, food web, and migration is discussed.相似文献
175.
Novel yeast protein kinase (YPK1 gene product) is a 40-kilodalton phosphotyrosyl protein associated with protein-tyrosine kinase activity. 总被引:23,自引:8,他引:15
下载免费PDF全文
![点击此处可从《Molecular and cellular biology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
D Dailey G L Schieven M Y Lim H Marquardt T Gilmore J Thorner G S Martin 《Molecular and cellular biology》1990,10(12):6244-6256
Extracts of bakers' yeast (Saccharomyces cerevisiae) contain protein-tyrosine kinase activity that can be detected with a synthetic Glu-Tyr copolymer as substrate (G. Schieven, J. Thorner, and G.S. Martin, Science 231:390-393, 1986). By using this assay in conjunction with ion-exchange and affinity chromatography, a soluble tyrosine kinase activity was purified over 8,000-fold from yeast extracts. The purified activity did not utilize typical substrates for mammalian protein-tyrosine kinases (enolase, casein, and histones). The level of tyrosine kinase activity at all steps of each preparation correlated with the content of a 40-kDa protein (p40). Upon incubation of the most highly purified fractions with Mn-ATP or Mg-ATP, p40 was the only protein phosphorylated on tyrosine. Immunoblotting of purified p40 or total yeast extracts with antiphosphotyrosine antibodies and phosphoamino acid analysis of 32P-labeled yeast proteins fractionated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated that the 40-kDa protein is normally phosphorylated at tyrosine in vivo. 32P-labeled p40 immunoprecipitated from extracts of metabolically labeled cells by affinity-purified anti-p40 antibodies contained both phosphoserine and phosphotyrosine. The gene encoding p40 (YPK1) was cloned from a yeast genomic library by using oligonucleotide probes designed on the basis of the sequence of purified peptides. As deduced from the nucleotide sequence of YPK1, p40 is homologous to known protein kinases, with features that resemble known protein-serine kinases more than known protein-tyrosine kinases. Thus, p40 is a protein kinase which is phosphorylated in vivo and in vitro at both tyrosine and serine residues; it may be a novel type of autophosphorylating tyrosine kinase, a bifunctional (serine/tyrosine-specific) protein kinase, or a serine kinase that is a substrate for an associated tyrosine kinase. 相似文献
176.
Identification of a 42-kilodalton phosphotyrosyl protein as a serine(threonine) protein kinase by renaturation. 总被引:17,自引:7,他引:10
下载免费PDF全文
![点击此处可从《Molecular and cellular biology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
We have surveyed fibroblast lysates for protein kinases that might be involved in mitogenesis. The assay we have used exploits the ability of blotted, sodium dodecyl sulfate-denatured proteins to regain enzymatic activity after guanidine treatment. About 20 electrophoretically distinct protein kinases could be detected by this method in lysates from NIH 3T3 cells. One of the kinases, a 42-kilodalton serine(threonine) kinase (PK42), was found to possess two- to fourfold-higher in vitro activity when isolated from serum-stimulated cells than when isolated from serum-starved cells. This kinase comigrated on sodium dodecyl sulfate-gels with a protein (p42) whose phosphotyrosine content increased in response to serum stimulation. The time courses of p42 tyrosine phosphorylation and PK42 activation were similar, reaching maximal levels within 10 min and returning to basal levels within 5 h. Both p42 tyrosine phosphorylation and PK42 activation were stimulated by low concentrations of phorbol esters, and the responses of p42 and PK42 to TPA were abolished by chronic 12-O-tetradecanoylphorbol-13-acetate (TPA) treatment. Chronic TPA treatment had less effect on serum-induced p42 tyrosine phosphorylation and PK42 activation. PK42 and p42 bound to DEAE-cellulose, and both eluted at a salt concentration of 250 mM. Thus, PK42 and p42 comigrate and cochromatograph, and the kinase activity of PK42 correlates with the tyrosine phosphorylation of p42. These findings suggest that PK42 and p42 are related or identical, that PK42 is activated by tyrosine phosphorylation, and that this tyrosine phosphorylation can be regulated by protein kinase C. 相似文献
177.
High-frequency disruption of the N-myc gene in embryonic stem and pre-B cell lines by homologous recombination. 总被引:15,自引:4,他引:11
下载免费PDF全文
![点击此处可从《Molecular and cellular biology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
J Charron B A Malynn E J Robertson S P Goff F W Alt 《Molecular and cellular biology》1990,10(4):1799-1804
Identification of gene function has often relied on isolation of mutant cells in which expression of the gene was inactivated. Gene targeting by homologous recombination in tissue culture now may provide a technology to rapidly and directly produce such mutant mammalian cells. We demonstrate that selection of embryonic stem and pre-B cell lines for expression of a promoterless construct containing murine N-myc genomic sequences fused to a gene encoding neomycin resistance allows highly efficient recovery of variants in which the endogenous N-myc gene is disrupted. The high frequency of N-myc gene disruption by this method should permit targeted disruption of both allelic N-myc copies in various cell lines to study N-myc function. 相似文献
178.
The pathway for the aerobic catabolism of 1,3,5-trihydroxybenzene (phloroglucinol) by a new strain of Penicillium was investigated using both in vivo and in vitro cell-free systems. The fungal strain was isolated by enrichment on phloroglucinol and identified as P. simplicissimum (Oud) Thom. It grew optimally at pH 5.5 and 27°C with 119 mM (1.5%w/v) of phloroglucinol in a basal mineral salts medium. Vapours of the crystalline substrate placed in a Petri-plate lid supported the growth of the fungal colonies on the agar surface. Mycelia grown on phloroglucinol accumulated 1,2,4-trihydroxybenzene and resorcinol in the medium. Washed, resting mycelia grown on phloroglucinol, when resuspended in a buffer utilized oxygen in the presence of catechol, resorcinol, pyrogallol and phloroglucinol. A NADPH-dependent reductase in the cell-free extract reduced phloroglucinol to dihydrophloroglucinol. This electron donor could not be replaced by NADH. Resorcinol hydroxylase, phloroglucinol reductase, catechol-1,2-oxygenase, and catechol-2,3-oxygenase were detected in cell-free extracts of mycelia grown on phloroglucinol. The possible steps in the degradation of phloroglucinol are discussed. 相似文献
179.
Martin Poot Julia Koehler Peter S. Rabinovitch Holger Hoehn Jean H. Priest 《Human genetics》1990,84(3):258-262
Summary BrdU-Hoechst flow cytometry was used to investigate the effects of DNA hypomethylation, induced by treatment with 5-azacytidine (5AC), on cell proliferation. When human fibroblast-like cells derived from skin and amniotic fluid were exposed to 5AC during three successive cell cycles, their clone-forming ability was diminished after removal of the drug. Treated cells were rendered quiescent by culture with low serum in the absence of the drug. Upon serum stimulation, they showed a diminished fraction of proliferating cells, which exhibited a prolonged transit through the S and G2 phase of the cell cycle, and a permanent arrest within the G2 compartment. This pattern of disturbed cell proliferation may in part explain the changes in replication banding pattern reported in the literature. Cytogenetic analysis of 5AC-treated cells revealed numerous endomitoses and tetraploid metaphases indicating a disturbed chromosome cycle in association with these cell kinetic perturbations. 相似文献
180.
Increased frequency of 6-thioguanine-resistant peripheral blood lymphocytes in Werner syndrome patients 总被引:1,自引:1,他引:0
Ken-ichiro Fukuchi Kiyoji Tanaka Yuichi Kumahara Kazuo Marumo Matthew B. Pride George M. Martin Raymond J. Monnat Jr. 《Human genetics》1990,84(3):249-252
Summary The frequency of spontaneous 6-thioguanine (TG)-resistant peripheral blood lymphocytes in five unrelated Werner syndrome (WS) patients was determined using an autoradiographic labeling assay. The average frequency of TG-resistant lymphocytes was eightfold higher in WS patients than in sex- and age-matched normal control donors. This finding and previous identification of increased spontaneous chromosomal rearrangements and deletions in WS cells or cell lines suggest that WS is a human genomic instability or mutator syndrome. 相似文献