全文获取类型
收费全文 | 6420篇 |
免费 | 471篇 |
国内免费 | 1篇 |
出版年
2023年 | 23篇 |
2022年 | 72篇 |
2021年 | 131篇 |
2020年 | 79篇 |
2019年 | 139篇 |
2018年 | 181篇 |
2017年 | 144篇 |
2016年 | 224篇 |
2015年 | 288篇 |
2014年 | 381篇 |
2013年 | 454篇 |
2012年 | 496篇 |
2011年 | 468篇 |
2010年 | 311篇 |
2009年 | 252篇 |
2008年 | 381篇 |
2007年 | 350篇 |
2006年 | 292篇 |
2005年 | 295篇 |
2004年 | 302篇 |
2003年 | 269篇 |
2002年 | 242篇 |
2001年 | 103篇 |
2000年 | 79篇 |
1999年 | 74篇 |
1998年 | 53篇 |
1997年 | 40篇 |
1996年 | 58篇 |
1995年 | 32篇 |
1994年 | 33篇 |
1993年 | 33篇 |
1992年 | 57篇 |
1991年 | 48篇 |
1990年 | 43篇 |
1989年 | 34篇 |
1988年 | 39篇 |
1987年 | 29篇 |
1986年 | 28篇 |
1985年 | 31篇 |
1984年 | 19篇 |
1983年 | 31篇 |
1982年 | 26篇 |
1981年 | 24篇 |
1980年 | 23篇 |
1979年 | 18篇 |
1977年 | 18篇 |
1976年 | 15篇 |
1975年 | 19篇 |
1974年 | 17篇 |
1972年 | 16篇 |
排序方式: 共有6892条查询结果,搜索用时 15 毫秒
31.
Rosa Sorrentino Carlo Iannicola Sandro Costanzi Alberto Chersi Roberto Tosi 《Immunogenetics》1991,33(2):118-123
DNA molecules derived from three alleles of the HLA-DRB3 locus and differing from each other at several nucleotide sites were denatured and cross-hybridized. Each allelic combination was found to generate a pair of heteroduplexes of different mobility. Their retardation as compared to homoduplexes was proportional to the number of mismatches. In each heteroduplexes pair the component possessing the highest number of Pyr-Pyr oppositions was the most retarded. The results are those predicted by a theoretical model implying a correlation between base-pair opening and bending of the DNA double helix. These observations introduce a new HLA typing method at the genomic level and indicate an experimental approach to the analysis of the superhelical DNA conformation as related to different types of base oppositions. 相似文献
32.
33.
Purification and characterization of aspartate aminotransferase from the thermoacidophilic archaebacterium Sulfolobus solfataricus 总被引:4,自引:0,他引:4
G Marino G Nitti M I Arnone G Sannia A Gambacorta M De Rosa 《The Journal of biological chemistry》1988,263(25):12305-12309
Aspartate aminotransferase from the archaebacterium Sulfolobus solfataricus, a thermoacidophilic organism isolated from an acidic hot spring (optimal growth conditions: 87 degrees C, pH 3.5) was purified to homogeneity. The enzyme is a dimer (Mr subunit = 53,000) showing microheterogeneity when submitted to chromatofocusing and/or isoelectric focusing analysis (two main bands having pI = 6.8 and 6.3 were observed). The N-terminal sequence (22 residues) does not show any homology with any stretch of known sequence of aspartate aminotransferases from animal and bacterial sources. The apoenzyme can be reconstituted with pyridoxamine 5'-phosphate and/or pyridoxal 5'-phosphate, each subunit binding 1 mol of coenzyme. The absorption maxima of the pyridoxamine and pyridoxal form are centered at 325 and 335 nm, respectively; the shape of the pyridoxal form band does not change with pH. The enzyme has an optimum temperature higher than 95 degrees C, and at 100 degrees C shows a half-inactivation time of 2 h. The above properties seem to be unique even for enzymes from extreme thermophiles (Daniel, R. M. (1986) in Protein Structure, Folding, and Design (Oxender, D. L., ed) pp. 291-296, Alan R. Liss, Inc., New York) and lead to the conclusion that aspartate aminotransferase from S. solfataricus is one of the most thermophilic and thermostable enzymes so far known. 相似文献
34.
Y Blouquit M D Rhoda J Delanoe-Garin R Rosa J C Prome C Poyart G Puzo J M Bernassau J Rosa 《The Journal of biological chemistry》1986,261(15):6758-6764
A new minor Hb fraction initially designated Hbx, has been found in the hemolysate of an erythremic patient that we have previously described with a complete erythrocyte bisphosphoglycerate mutase (EC 5.4.2.4) deficiency. Hbx (3.5% of the total) was detected by isoelectric focusing and exhibited electrophoretic and chromatographic properties similar to those of several variants of the Hb central cavity. By density fractionation of red cells, it was demonstrated that Hbx was an aging hemoglobin as in the case of glycated Hb A1c. Functional studies revealed a low oxygen affinity and almost complete inhibition of the allosteric effect of the organic phosphate effectors. Structural studies demonstrated an absence of tryptic cleavage between the peptides beta T9 and beta T10 suggesting the presence of an adduct on Lys beta 82 or on a neighboring residue. Fast atom bombardment mass spectrometry and a specific enzymatic assay with glyoxylate reductase demonstrated that the beta 82 adduct was a glycerate moiety. It was concluded that Hbx was a glycerylated Hb, alpha 2A beta 2(82) (EF6) N epsilon-glyceryllysine, to our knowledge the first example of glycerylated protein. The mechanism of formation of glyceryl Hb, which was found in the four studied subjects with a bisphosphoglyceromutase deficiency, remains to be determined. 相似文献
35.
G Pontoni C Manna A Salluzzo L del Piano P Galletti M De Rosa V Zappia 《Biochimica et biophysica acta》1985,836(2):222-232
In order to elucidate the reaction mechanism and the substrate-binding sites, CDPcholine:1,2-diacylglycerol cholinephosphotransferase (EC 2.7.8.2), prepared from rat liver microsomal fraction, has been subjected to kinetic analysis and substrate specificity studies. Kinetic evidence supports the hypothesis of a Bi-Bi sequential mechanism, involving a direct nucleophilic attack of diacylglycerol on CDPcholine during the reaction. To investigate the substrate requirements for recognition and catalysis, several CDPcholine analogs, modified in the nitrogen base or in the sugar or in the pyrophosphate bridge, have been synthesized, characterized and assayed as substrates and/or inhibitors of the reaction. The amino group on the pyrimidine ring, the 2'-alcoholic function of the ribose moiety as well as the pyrophosphate bridge have been identified as critical sites for enzyme-substrates interactions. 相似文献
36.
Annette Gulik Vittorio Luzzati Mario De Rosa Agata Gambacorta 《Journal of molecular biology》1985,182(1):131-149
We describe in this work the structure and polymorphism of a variety of lipids extracted from Sulfolobus solfataricus, an extreme thermoacidophilic archaebacterium growing at about 85 °C and pH 2. These lipids are quite different from the usual fatty acid lipids of eukaryotes and prokaryotes: each molecule consists of two C40 ω-ω′ biphytanyl residues (with 0 to 4 cyclopentane groups per residue), ether linked at both ends to two (variably substituted) glycerol or nonitol groups. Four lipid preparations were studied; the total and the polar lipid extracts, and two hydrolytic fractions, the symmetric glycerol dialkyl glycerol tetraether and the asymmetric glycerol dialkyl nonitol tetraether, as a function of water content and temperature, using X-ray scattering techniques. The main conclusions from the study of the four lipid preparations can be summarized as follows. (1) As with other lipids, a remarkable number and variety of phases are observed over a temperature-concentration range close to “physiological” conditions. The possibility is discussed that this polymorphism reflects a fundamental property of lipids, closely related to their physiological rôle. (2) As in other lipids, two types of chain conformations are observed: a disordered one (type α) at high temperature; at lower temperature, a more ordered packing of stiff chains, all parallel to each other (type β′). At temperatures and degrees of hydration approaching the conditions prevailing in the living cell, the conformation is of type α. (3) In all the phases with chains in the α conformation, the unsubstituted glycerol headgroups, whose concentration is high in these lipids, segregate in the hydrocarbon matrix, away from the other polar groups. This property may have interesting biological consequences: for example, the chains of a fraction of the bipolar lipid molecules can span hydrocarbon gaps as wide as 75 Å. (4) Two cubic phases are observed in the total and the polar lipid extracts, which display a remarkable degree of metastability, most unusual in lipid phase transitions involving structures with chains in the α conformation. This phenomenon can be explained by the interplay of the physical structure of the cubic phases (the two contain two intertwined and unconnected three-dimensional networks of rods) and the chemical structure of the lipid molecules: the two headgroups of most molecules being anchored on each of the two networks of rods, the migration of the lipid molecules is hindered by the two independent diffusion processes and by the entanglement of the chains. The possibility is discussed that this phenomenon may reflect an evolutionary response to a challenge of the natural habitat of these archaebacteria. 相似文献
37.
W. L. C. Vaz D. Hallmann R. M. Clegg A. Gambacorta M. De Rosa 《European biophysics journal : EBJ》1985,12(1):19-24
We have used the fluorescence recovery after photobleaching technique to study the translational diffusion, in L phase multibilayers of 1-palmitoyl-2-oleoylphosphatidylcholine (POPC), of fluorescent derivatives of 1-palmitoyl-2-oleoylphosphatidylethanolamine (NBD-POPE) and a membrane-spanning phosphatidylethanolamine (NBD-MSPE). The latter derivative was prepared from a membrane-spanning glycerol-dialkyl-glycerol tetraether lipid isolated from the thermophilic and acidophilic archaebacterium Sulfolobus solfataricus. The translational diffusion was examined between about 15° and 45°C. It is shown that over this temperature range the translational diffusion coefficient for NBD-MSPE is 2/3 that for NBD-POPE which spans only one monolayer of the bilayer. The result is interpreted in terms of existing models for translational diffusion in lipid membranes.Abbreviations D
t
translational diffusion coefficient
- FRAP
fluorescence recovery after photobleaching
- MSPE
a membrane-spanning phosphatidylethanolamine derived from a glycerol-dialkyl-glycerol tetraether lipid isolated from Sulfolobus solfataricus
- NBD
4-nitrobenz-2-oxa-1,3-diazolyl
- PE
phosphatidylethanolamine
- POPC
1-palmitoyl-2-oleoylphosphatidylcholine
- POPE
1-palmitoyl-2-oleoylphosphatidylethanolamine 相似文献
38.
S. Bruno S. Cannistraro A. Gliozzi M. De Rosa A. Gambacorta 《European biophysics journal : EBJ》1985,13(2):67-76
A spin label study has been carried out on bipolar lipids extracted from Sulfolobus solfataricus, an extreme thermophilic archaebacterium growing at about 85°C and pH 3. These lipids are cyclic diisopranyl tetraether molecules, quite different from the usual fatty acid lipids. Two hydrolytic fractions of the membrane complex lipids have been studied: the symmetric lipid glycerol-dialkyl-glycerol-tetraether (GDGT) and the asymmetric lipid glyceroldialkyl-nonitol-tetraether (GDNT). The ESR spectra confirm the results previously obtained from calorimetric and X-ray diffraction experiments showing a polymorphic behaviour of these lipids and indicating the critical temperature ranges at which structural transitions occur. Moreover, the present study adds information on the dynamics of the different portions of the hydrophobic chain. ST-ESR measurements show correlation times ranging from 10-8 s up to 10-5 s, depending upon the lipid sample, the label position and the degree of hydration. At very high temperatures, i.e. the physiological temperatures of Sulfolobus solfataricus, the nonitol head groups of the asymmetric lipids form a strongly immobilized structure. Indeed, the molecular correlation times of the outermost hydrophobic portion of GDNT are higher, by a factor up to 103, than those of usual monopolar lipids. Anisotropic motional behaviour is observed even at such very high temperatures. Possible biological implications are discussed.Abbreviations used are ESR
electron spin resonance
- St-ESR
saturation transfer electron spin resonance
- GDGT
glyceroldialkyl-glycerol-tetracther
- GDNT
glycerol-dialkyl-nonitoltetraether
- 5 SASL
12SASL and 16SASL, stearic acid spin labels, N-oxyl-4,4-dimethyloxazolidine derivatives of 5-ketostearic acid, 12-ketostearic acid and 16-ketostearic acid, respectively
- DSC
differential scanning calorimetry 相似文献
39.
Evidence for a rate-limiting role of cysteinesulfinate decarboxylase activity in taurine biosynthesis in vivo 总被引:1,自引:0,他引:1
J de la Rosa M H Stipanuk 《Comparative biochemistry and physiology. B, Comparative biochemistry》1985,81(3):565-571
The relationship between activities of enzymes involved in cysteine oxidation and the apparent conversion of cysteine to taurine in vivo were investigated in the rat and cat. Both hepatic cysteinesulfinate decarboxylase activity and the oxidation in vivo of cysteine to taurine were lower in the kitten than in the adult female rat and lower in the latter than in the young male rat. Our data support the hypothesis that cysteinesulfinate decarboxylase plays a rate-limiting role in taurine biosynthesis. 相似文献
40.
A galactofuranosyl-containing glycopeptide has been isolated from mycelium ofAscobolus furfuraceus by extraction with water. The glycoconjugate was purified by DEAE-cellulose chromatography followed by gel filtration. A molecular weight of about 20 000 was determined by the latter method using standard dextrans. Neutral sugars accounted for 94.5% of the glycopeptide and were characterized as mannose, galactose, and glucose. Glucosamine was estimated colorimetrically (1.8%). The molar ratio of Man:Gal:Glc:GlcNH2 was 68:32:16:2. A trace amount of total phosphorus (0.2%) was found. The predominant amino acids were threonine and serine. The peptide moiety was labeled with [14C]formaldehyde and the elution of radioactivity was coincident with sugar on gel filtration in the presence of sodium dodecyl sulfate. The peak of radioactivity was retarded on release of galactose by mild acid hydrolysis. These results confirm the sugar-peptide linkage. 相似文献