全文获取类型
收费全文 | 6076篇 |
免费 | 189篇 |
国内免费 | 316篇 |
出版年
2023年 | 45篇 |
2022年 | 84篇 |
2021年 | 105篇 |
2020年 | 122篇 |
2019年 | 181篇 |
2018年 | 158篇 |
2017年 | 109篇 |
2016年 | 159篇 |
2015年 | 174篇 |
2014年 | 364篇 |
2013年 | 494篇 |
2012年 | 185篇 |
2011年 | 354篇 |
2010年 | 285篇 |
2009年 | 335篇 |
2008年 | 377篇 |
2007年 | 364篇 |
2006年 | 314篇 |
2005年 | 294篇 |
2004年 | 197篇 |
2003年 | 218篇 |
2002年 | 210篇 |
2001年 | 103篇 |
2000年 | 96篇 |
1999年 | 103篇 |
1998年 | 91篇 |
1997年 | 77篇 |
1996年 | 65篇 |
1995年 | 74篇 |
1994年 | 83篇 |
1993年 | 73篇 |
1992年 | 62篇 |
1991年 | 48篇 |
1990年 | 44篇 |
1989年 | 47篇 |
1988年 | 39篇 |
1987年 | 35篇 |
1986年 | 26篇 |
1985年 | 56篇 |
1984年 | 102篇 |
1983年 | 72篇 |
1982年 | 38篇 |
1981年 | 39篇 |
1980年 | 28篇 |
1979年 | 10篇 |
1978年 | 14篇 |
1977年 | 7篇 |
1976年 | 9篇 |
1975年 | 5篇 |
1973年 | 3篇 |
排序方式: 共有6581条查询结果,搜索用时 15 毫秒
91.
Ken Lee Akihiro Ito †Kunio Koshimura †Tetsuya Ohue †Yasutaka Takagi †Soichi Miwa 《Journal of neurochemistry》1995,64(2):874-882
Abstract: Hypoxia is known to disturb neuronal signal transmission at the synapse. Presynaptically, hypoxia is reported to suppress the release of neurotransmitters, but its postsynaptic effects, especially on the function of neurotransmitter receptors, have not yet been elucidated. To clarify the postsynaptic effects, we used cultured bovine adrenal chromaffin cells as a model of postsynaptic neurons and examined specific binding of l -[3 H]nicotine (an agonist for nicotinic acetylcholine receptors: nAChRs) and 22 Na+ flux under control and hypoxic conditions. Experiments were performed in media preequilibrated with a gas mixture of either 21% O2 /79% N2 (control) or 100% N2 (hypoxia). Scatchard analysis of the specific binding to the cells revealed that the KD under hypoxic conditions was twice as large as that under control conditions, whereas the B max was unchanged. When the specific [3 H]nicotine binding was kinetically analyzed, the association constant ( k 1 ) but not the dissociation constant ( k −1 ) was decreased to 40% of the control value by hypoxia. When the binding assay was performed using the membrane fraction, these changes were not observed. Nicotine-evoked 22 Na+ flux into the cells was suppressed by hypoxia. In contrast, specific [3 H]quinuclidinyl benzilate binding to the intact cells was unaffected by hypoxia. These results demonstrate that hypoxia specifically suppresses the function of nAChRs (and hence, neuronal signal transmission through nAChRs), primarily by acting intracellularly. 相似文献
92.
Craig J. Coates Catherine L. Turney Marianne Frommer David A. O'Brochta W. D. Warren Peter W. Atkinson 《Molecular & general genetics : MGG》1995,249(2):246-252
Plasmid-based excision assays performed in embryos of two non-drosophilid species using the mariner transposable element from Drosophila mauritiana resulted in empty excision sites identical to those observed after the excision of mariner from D. mauritiana chromosomes. In the presence of the autonomous mariner element Mos1, excision products were recovered from D. melanogaster, D. mauritiana and the blowfly Lucilia cuprina. When a hsp82 heat shock promoter-Mos1 construct was used to supply mariner transposase, excision products were also recovered from the Queensland fruitfly Bactrocera tryoni. Analysis of DNA sequences at empty excision sites led us to hypothesise that the mariner excision/repair process involves the formation of a heteroduplex at the excision breakpoint. The success of these assays suggests that they will provide a valuable tool for assessing the ability of mariner and mariner-like elements to function in non-drosophilid insects and for investigating the basic mechanisms of mariner excision and repair. 相似文献
93.
Expression of betaine aldehyde dehydrogenase gene and salinity tolerance in rice transgenic plants 总被引:9,自引:0,他引:9
Betaine as one of osmolytes plays an important role in osmoregulation of most high plants. Betaine aldehyde dehydrogenase C BADH) is the second enzyme involved in betaine biosynthesis. The BADH gene from a halophite, Atriplex hortensis, was transformed into rice cultivars by bombarment method. Totally 192 transgenic rice plants were obtained and most of them had higher salt tolerance than controls. Among transgenic plants transplanted in the saline pool containing 0.5% NaCl in a greenhouse, 22 survived, 13 of which set seeds, and the frequency of seed setting was very low, only 10% . But the controls could not grow under the same condition. The results of BADH ac-tivity assay and Northern blot showed that the BADH gene was integrated into chromosomes of transgenic plants and expressed. 相似文献
94.
高必需氨基酸转基因马铃薯的研究 总被引:8,自引:0,他引:8
80年代以来,马铃薯遗传转化系统日趋成熟,转基因工程植株已被广泛应用于基础科学研究[1]。作为食物蛋白和能量主要来源的马铃薯,提高其蛋白质含量及质量的遗传工程研究正受到人们的普遍关注[2]。Yang等[2]将旨在改善氨基酸平衡的CAT-HEAAE(氯酶素乙酰转移酶-高含量人体必需氨基酸)融合基因导入马铃薯,获得了Southernblot、Northernblot、Westernblot的证据,但尚缺少氨基酸分析的资料。玉米醇溶蛋白(zein)[3]是一个富含甲硫氨酸的贮存蛋白,它和人工合成的HE… 相似文献
95.
96.
Rifampin and chloramphenicol inhibited the synthesis of collagenase of Streptomyces sp. A8, suggesting de novo synthesis. The collagenase was induced by insoluble collagen, its macromolecular fragments, gelatin, peptone, hide powder and yeast extract. Growth as well as collagenase synthesis were dependent on substrate availability. Purification of collagenase by DEAE-cellulose chromatography resulted in approximately 25-fold increase in activity (268.6 μmol glycine equivalents min?1 mg?1 protein) relative to the activity of the culture filtrate (10.5 μmol glycine equivalents min?1 mg?1 protein). 相似文献
97.
The state of the art of firefly luciferase research is reviewed with special emphasis on its purification and immobilization. The notion of bioluminescence and its role in APT monitoring is described. The need to purify luciferase and the advantages of immobilization are discussed. An insight into the existing methods of luciferase purification and immobilization is given. The scope of the bioluminescent assay is underlined. 相似文献
98.
The binding of monoiodo 125I-Trp11-neurotensin to purified rat gastric fundus smooth muscle plasma membranes was characterized. Specific binding of ligand in subcellular fractions from rat fundus smooth muscle showed a distribution that paralleled that of several plasma membrane marker enzymes. 125I-Trp11-neurotensin binding to smooth muscle plasma membranes at 25 degrees C was maximal at 30 min, reversible and saturable. Scatchard analysis of equilibrium data indicated the existence of two classes of binding sites with dissociation constants (Kd) of 56 pmol and 1.92 nM, and corresponding binding capacities (Bmax) of 6.6 fmol/mg and 11.4 fmol/mg of membrane protein. Analogues and fragments of neurotensin competed for 125I-Trp11-neurotensin binding with a rank order of potency similar to that previously reported for their contracting effect in rat fundus strips. Na+ decreased in a concentration dependent manner the binding of labelled ligand to the high affinity site. At 100 mM, Na+ induced a 6-fold increase in the IC50 of neurotensin for inhibition of 125I-Trp11-neurotensin binding. At this concentration of Na+, the IC50 for neurotensin was 1 nM, a value close to the Kd of the low affinity site. 相似文献
99.
Georg R. Beilharz Carl R. Middlehurst Philip W. Kuchel Glenn E. Hunt Gordon F.S. Johnson 《Analytical biochemistry》1984,137(2):324-329
Detailed operating conditions are reported for the determination of choline in human erythrocytes using proton nuclear magnetic resonance spectroscopy in conjunction with the inversion-recovery spin-echo pulse sequence. The results of the NMR method were in excellent agreement with those obtained using an enzymatic (choline oxidase) assay; however, they were approximately three times higher than those reported using gas chromatography/mass spectrometry techniques. The differences may be partly due to the method of preparing or sampling cells since there is a distribution of choline in cells of different ages. However, choline levels were not affected by the methods used in the present study for storing or preparing cells. 相似文献
100.
A fluorogenic peptide, dansyl-D-Ala-Gly-Phe(pNO2)-Gly (DAGNPG), was synthesized as a selective substrate for the neutral metalloendopeptidase (EC 3.4.24.11) involved in enkephalin metabolism. This enzyme, designated "enkephalinase," cleaves the Gly-Phe(pNO2) peptide bond of DAGNPG (V = 0.65 mumol/mg protein/min and Km = 45 microM) leading to a fluorescence increase related to the disappearance of intramolecular quenching of the dansyl fluorescence by the nitrophenyl residue. This change was used for quantitative measurements of "enkephalinase" activity in different tissues and determination of inhibitory potency of various compounds. The substrate is not cleaved by aminopeptidase or dipeptidylaminopeptidase activities and the assay itself is rapid, convenient, and sensitive. 相似文献