全文获取类型
收费全文 | 31232篇 |
免费 | 2441篇 |
国内免费 | 1787篇 |
出版年
2024年 | 59篇 |
2023年 | 325篇 |
2022年 | 849篇 |
2021年 | 1401篇 |
2020年 | 962篇 |
2019年 | 1129篇 |
2018年 | 1189篇 |
2017年 | 869篇 |
2016年 | 1308篇 |
2015年 | 1792篇 |
2014年 | 2063篇 |
2013年 | 2363篇 |
2012年 | 2748篇 |
2011年 | 2490篇 |
2010年 | 1524篇 |
2009年 | 1343篇 |
2008年 | 1624篇 |
2007年 | 1478篇 |
2006年 | 1332篇 |
2005年 | 1165篇 |
2004年 | 1058篇 |
2003年 | 967篇 |
2002年 | 872篇 |
2001年 | 541篇 |
2000年 | 470篇 |
1999年 | 462篇 |
1998年 | 325篇 |
1997年 | 328篇 |
1996年 | 318篇 |
1995年 | 299篇 |
1994年 | 252篇 |
1993年 | 182篇 |
1992年 | 254篇 |
1991年 | 175篇 |
1990年 | 166篇 |
1989年 | 127篇 |
1988年 | 86篇 |
1987年 | 109篇 |
1986年 | 74篇 |
1985年 | 81篇 |
1984年 | 69篇 |
1983年 | 40篇 |
1982年 | 45篇 |
1981年 | 35篇 |
1980年 | 22篇 |
1979年 | 15篇 |
1978年 | 12篇 |
1977年 | 9篇 |
1976年 | 10篇 |
1975年 | 11篇 |
排序方式: 共有10000条查询结果,搜索用时 15 毫秒
51.
52.
53.
S. Honecker B. Bisping Zhu Yang H. -J. Rehm 《Applied microbiology and biotechnology》1989,31(1):17-24
Summary Immobilized cells of Aspergillus niger needed a lower initial sucrose concentration than free cells in order to obtain maximal yields of citric acid production. High sucrose concentrations led to reduced yields and increased polyol formation (glycerol, erythritol, arabitol). Continuous fermentation with media containing low sugar concentrations prevented the formation of polyols. The change from nitrogen-limited to phosphate-limited precultivation of immobilized spores significantly increased the productivity of the mycelium. The ratio of citric acid to residual sugar in the effluent distinctly lay in the direction of citric acid. Inside the alginate beads mainly large bulbous cells were observed. 相似文献
54.
P Favard N Favard Q L Zhu J Bourguet J P Lechaire 《Biology of the cell / under the auspices of the European Cell Biology Organization》1989,66(1-2):99-106
We have developed a technique for recovering apical membranous sheets from amphibian urinary bladders by gelatin stripping. The tissue is mounted on a lucite support and the apical surface is first stuck onto a gelatin-coated glass slide at 30 degrees C. This sandwich is then chilled on ice and the bladder is pulled away from the slide. Preliminary results indicate that this simple technique could be used to remove membranous apical sheets of various sizes, almost devoid of cytoplasmic contamination and without significant damage to the underlying cell structures. The method could also be adapted to prepare perforated cells and to study the cohesive forces between the different layers of the tissue. 相似文献
55.
Teresa Urbanik-Sypniewska Ulrich Seydel Michaela Greck Jürgen Weckesser Hubert Mayer 《Archives of microbiology》1989,152(6):527-532
The structure of the lipopolysaccharide from Rhizobium meliloti 10406, a derivative of the wild-type strain MVII-1, was examined. The compositional analysis of its polysaccharide moiety demonstrated lack of heptose(s), but high contents in glucose, galacturonic acid and 2-keto-3-deoxy-octonate (dOclA) as characteristic features. The lipid A moiety consisted of a -1,6 linked glucosamine disaccharide carrying ester (at C-4) and glycosidically (at C-1) linked phosphate residues, both present exclusively as monoester phosphates but not as phosphodiesters. Ester- and amidelinked 3-hydroxy fatty acids were mostly present as non-3-O-acylated residues. Laser desorption mass spectrometry (LD-MS) revealed heterogeneity in the fatty acid substitution, as was also indicated by the non-stoichiometric ratios obtained by quantitative fatty acid analysis. The predominating lipid A structure contained at the reducing glucosamine residue ester-linked 3-hydroxy-tetradecanoic acid (3-OH-14:0) and amide-linked 3-OH-18:0, or 3-OH-18:1, respectively. The distal (non-reducing) glucosamine carried ester-bound the recently discovered 27-hydroxyoctacosanoic acid and 3-OH-14:0 and, as amide-linked fatty acid, mostly 3-hydroxy-stearic acid (3-OH-18:0).The isolated lipopolysaccharide exhibited a high extent of lethal toxicity in galactosamine-treated mice, comparable to that of enterobacterial lipopolysaccharide. The structural relationship of LPS and lipid A of Rhizobium meliloti to other rhizobial lipopolysaccharides and lipid A's with respect to questions of taxonomy and of phylogenetic relationships will be discussed.Abbreviations LPS
lipopolysaccharide
- dOclA
3-deoxy-D-mannooctulosonic acid (KDO)
- GalA
galacturonic acid
- DOC
sodium deoxycholate
- PAGE
polyacrylamide gel electrophoresis
- LD-MS
laser desorption-mass spectrometry 相似文献
56.
D L Zhu 《Analytical biochemistry》1989,177(1):120-124
A simple and efficient site-specific mutagenesis method is described. First, a single-stranded (ss) circular vector is linearized at the site where the desired mutation will be introduced. To do this, an oligodeoxynucleotide complementary to the target region of the ss vector and containing a restriction enzyme recognition sequence is annealed to the circular ss vector, and the partial double-strand formed is subsequently cleaved with that enzyme. Then, another oligodeoxynucleotide spanning the nick and carrying the mutation is annealed to the linearized ss DNA template and the annealed mixture is used directly to transform Escherichia coli without prior enzymatic DNA synthesis in vitro. The procedure has been applied successfully to constructing insertion, deletion, and point mutations in both M13 phage vectors and plasmid vectors containing the f1 origin of replication. 相似文献
57.
G C Chen D A Hardman R L Hamilton C M Mendel J W Schilling S Zhu K Lau J S Wong J P Kane 《Biochemistry》1989,28(6):2477-2484
The distribution of lipid-binding regions of human apolipoprotein B-100 has been investigated by recombining proteolytic fragments of B-100 with lipids and characterizing the lipid-bound fragments by peptide mapping, amino acid sequencing, and immunoblotting. Fragments of B-100 were generated by digestion of low-density lipoproteins (LDL) in the presence of sodium decyl sulfate with either Staphylococcus aureus V8 protease, pancreatic elastase, or chymotrypsin. Particles with electron microscopic appearance of native lipoproteins formed spontaneously when detergent was removed by dialysis from enzyme digests containing fragments of B-100 and endogenous lipids, or from incubation mixtures of delipidated B-100 fragments mixed with microemulsions of exogenous lipids (cholesteryl oleate and egg phosphatidylcholine). Fractionation of the recombinant particles by isopycnic or density gradient ultracentrifugation yielded complexes similar to native LDL with respect to shape, diameter, electrophoretic mobility, and surface and core compositions. Circular dichroic spectra of these particles showed helicity similar to LDL but a somewhat decreased content of beta-structure. Most of the fragments of B-100 were capable of binding to lipids; 12 were identified by direct sequence analysis and 14 by reaction with antisera against specific sequences within B-100. Our results indicate that lipid-binding regions of B-100 are widely distributed within the protein molecule and that proteolytic fragments derived from B-100 can reassociate in vitro with lipids to form LDL-like particles. 相似文献
58.
59.
Teresa Duda Stephen Chalberg Rameshwar K. Sharma 《Molecular and cellular biochemistry》1990,92(1):69-75
Summary A complementary DNA (cDNA) clone - cA2-47 - corresponding to a new 2-adrenergic receptor subtype has been isolated from a rat brain cDNA library and used as a hybridization probe to scrutinize the 2-receptor poly(A+) RNAs in rat brain, heart and adrenal gland. Hybridization of the 5 half of the coding region of this cDNA at 37°C to rat brain poly(A+) RNA revealed a single band at 5.8 kb as the size of its corresponding mRNA. Under identical hybridization conditions, a human platelet 2-receptor genomic probe failed to hybridize to any rat brain mRNAs.Under lower stringency conditions, hybridization of the full-length cDNA, cA2-47, to selected rat tissue poly(A+) RNA showed the presence of four different sized mRNAs in brain and three in both heart and adrenal gland. Messages of 1.3 kb and 2.1 kb were common in all three tissues (although the band at 2.1 kb was slightly higher in the heart and adrenal gland). A 5.8 kb mRNA was unique to the brain and a slightly higher band at 6.0 kb was consistently present in heart and adrenal gland but was absent in the brain. A fourth message at 3.4 kb was found predominantly in the brain and was either absent or present at very low levels in the other tissues examined. Under the same conditions, a human platelet 2-receptor probe hybridized to similar sized messages of 2.1 and 5.8 kb in rat brain and 2.2 and 6.0 kb in rat heart and adrenal gland. This probe, however, failed to detect the abundant 1.3 kb mRNA common to all tissues or the 3.4 kb message in rat brain. The extent of homology of these messages with cA2-47 is not confined to limited regions of the cDNA since similar hybridization patterns were observed using either 5-noncoding or 5-coding regions of the probe.These results provide the first direct evidence of a surprisingly large range of mRNA sizes for members of the 2-receptor family in brain, heart, and adrenal gland. The unique nature of certain members of the family in each of the tissues examined raises the curious possibility that these members might contribute to some of the individualized functions of the brain, cardiovasculature and adrenal gland. 相似文献
60.
Dale D. Brown Ronald G. Knowlton Joseph Hamill Teresa L. Schneider Ronald K. Hetzler 《European journal of applied physiology and occupational physiology》1990,60(3):179-182
The purpose of this study was to compare the physiological and biomechanical responses of wheelchair-dependent persons (WCD) to able-bodied persons (AB) during manual wheelchair ergometry. Five WCD and five AB performed a discontinuous wheelchair ergometer test starting at 12.8 W at 30 rev.min-1 (57 m.min-1) with increments of 7.0 W at 6-min intervals. Biomechanical data were collected 3.5 min into each stage followed by the collection of physiological data. After the fifth stage, peak oxygen consumption was determined by having the subject work against a resistance of 14.7-19.6 N at 30 rev.min-1. The WCD had significantly higher net mechanical efficiency at 26.7, 33.6 and 40.6 W in comparison to the AB. The WCD had significantly greater shoulder extension at the point of initial wheel contact as measured by the shoulder angle, while the AB had significantly greater shoulder range of motion at all work rates in comparison to the WCD. The results demonstrate that a significant physiological difference exists in the manner by which WCD and AB accomplish wheelchair ergometry. The biomechanical differences between AB and WCD were found to be a prominent factor contributing to the higher mechanical efficiency of WCD over AB. It was concluded that basic physiological and biomechanical differences exist between WCD and AB in manual wheelchair locomotion and that these differences are important considerations to the interpretation of data in wheelchair ergometry studies. 相似文献