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81.
Wolf-Dietrich Hard Jens M. Warnecke Roland K. Hartmann 《Molecular biology reports》1995,22(2-3):161-169
Modification interference is a powerful method to identify important functional groups in RNA molecules. We review here recent developments of techniques to screen for chemical modifications that interfere with (i) binding of(pre-)tRNA to bacterial RNase P RNA or (ii) pre-tRNA cleavage by this ribozyme. For example, two studies have analyzed positions at which a substitution of sulfur for thepro-Rp oxygen affects tRNA binding [1] or catalysis [2]. The results emphasize the functional key role of a central core element present in all known RNase P RNA subunits. The four sulfur substitutions identified in one study [2] to inhibit the catalytic step also interfered with binding of tRNA toE. coli RNase P RNA [1]. This suggests that losses in binding energy due to the modification at these positions affect the enzyme-substrate and the enzyme-transition state complex. In addition, the two studies have revealed, for the first time, sites of direct metal ion coordination in RNase P RNA. The potentials, limitations and interpretational ambiguities of modification interference experiments as well as factors influencing their outcome are discussed.Abbreviations nt
nucleotide(s)
- PAGE
polyacrylamide gel electrophoresis 相似文献
82.
Jens Meyer Norbert Elsner 《Journal of comparative physiology. A, Neuroethology, sensory, neural, and behavioral physiology》1995,176(4):563-573
1. | Up to 9 kHz, the tympanal membrane of the grasshopper Chorthippus biguttulus responds with equal sensitivity at the attachment sites of the low and the high-frequency receptors; at the latter site it is also particularly sensitive between 10 and 20 kHz. |
2. | The frequency spectra of the songs of both sexes exhibit maxima at 7–8 kHz, to which the membrane is well matched. In the high-frequency region, where the male songs have a peak at 30 kHz, there is no corresponding maximum in the membrane oscillation. |
3. | Because the tympanal membrane is immediately adjacent to air sacs in the tracheal system, it is deflected inward and outward by as much as 80 m during the respiratory cycle. |
4. | Measurements by laser vibrometry show that acoustically induced membrane oscillations are attenuated severely due to the respiratory displacement of the membrane for frequencies up to 10–12 kHz. By contrast, at higher frequencies the membrane sensitivity is doubled or tripled. |
5. | As a result of these membrane effects, the discharge in the tympanal nerve was profoundly reduced in the low-frequency range, whereas above 11 kHz there was a marked increase. This modulation of auditory sensitivity affects the animals' ability to detect conspecific songs. |
83.
Ethanol utilization regulatory protein: profile alignments give no evidence of origin through aldehyde and alcohol dehydrogenase gene fusion. 下载免费PDF全文
H. B. Nicholas Jr B. Persson H. Jrnvall J. Hempel 《Protein science : a publication of the Protein Society》1995,4(12):2621-2624
The suggestion that the ethanol regulatory protein from Aspergillus has its evolutionary origin in a gene fusion between aldehyde and alcohol dehydrogenase genes (Hawkins AR, Lamb HK, Radford A, Moore JD, 1994, Gene 146:145-158) has been tested by profile analysis with aldehyde and alcohol dehydrogenase family profiles. We show that the degree and kind of similarity observed between these profiles and the ethanol regulatory protein sequence is that expected from random sequences of the same composition. This level of similarity fails to support the suggested gene fusion. 相似文献
84.
Sulphur-heterotrophic growth exhibited a dual response to the expression of sulphate-assimilating enzymes. The level of ATP-sulphurylase (EC 2.7.7.4) appeared repressed while sulphite reductase (EC 1.8.7.1) and O-acetyl-l-serine sulphhydrylase (EC 4.2.99.8) were derepressed and coordinated in their occurrence. The capability of the cells to reduce adenylylphosphosulphate or 3-phospho adenylylphosphosulphate to cysteine coincided with the activity of sulphite reductase. The expression of these reducing steps lacked correlation with the regulation of ATP-sulphurylase.Abbreviations APS
adenylylphosphosulphate
- MVH
reduced methylviologen
- OAS
O-acetyl-l-serine
- PAPS
3-phospho adenylylphosulphate 相似文献
85.
An improved timm sulphide silver method for light and electron microscopic localization of heavy metals in biological tissues 总被引:3,自引:0,他引:3
Summary Modifications of the Timm sulphide silver method for the demonstration of heavy metals are described.To improve the structural preservation of the tissues perfusion with a glutaraldehyde fixative is employed before perfusion with the sodium sulphide solution. For the subsequent staining for light and electron microscopy, procedures for plastic embedding, paraffin embedding and cryostat sectioning are presented. Examples from several tissues are shown, including the pituitary, pancreas, intestine, tongue, kidney, testis and brain. The staining of autolytic, postmortal human brain tissue is demonstrated. 相似文献
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88.
Purification and characterization of an early glycoprotein from adenovirus type 2-infected cells. 下载免费PDF全文
An adenovirus type 2 early glycoprotein with an apparent molecular weight of 19,000 (E19K) in sodium dodecyl sulfate-polyacrylamide gels has been extensively purified. Purification involved detergent solubilization of membrane fractions from infected cells, followed by affinity chromatography on a lectin column and DEAE-Sephadex chromatography. The purified material contained three polypeptides (E40K, E19K, E17.5K), with approximately 90% of the material in the E19K moiety. All three polypeptides yielded identical tryptic peptide maps. The E19K polypeptide contained glucosamine as revealed by [3H]glucosamine labeling of infected cells and amino acid analysis of the purified protein. Immunoprecipitation with a monospecific antiserum showed that the E19K polypeptide started to be synthesized at 2 h, with a maximal rate at 4 h after infection. It was also synthesized at a low rate late in the infectious cycle (12 to 24 h postinfection). Immunoprecipitation from three adenovirus type 2-transformed hamster embryo cell lines and two adenovirus type 2-transformed rat cell lines revealed that one of the hamster cell lines (ad2HE4) and one of the rat cell lines (A2T2C4) expressed this protein. 相似文献
89.
90.
Stabursvik (1959) described the saponin fraction of Narthecium ossifragum as a sarsasapogenin glycoside with the structure arabinosegalactose-xylose-glucose-sarsasapogenin. In a renewed study of the phototoxic lamb disease alveld, in which this saponin has been implicated (Ender 1955), we have looked more closely at the saponin fraction. We find that there are two saponins, one major and one minor. Both have a branched trisaccharide on C-3 of the sapogenin. Galactose is directly attached to C-3 in both saponins. The major saponin has glucose and arabinose attached to galactose, the minor saponin has glucose and xylose. We suggest the names narthecin and xylosin for the spirostanol form of these two saponins. In fresh juice from leaves we find little narthecin, however. Most of the saponin is present in the furostanol form, with glucose on C-26. Enzymatic hydrolysis showed this glucose to be bound as a β-glucoside. From specific rotations in partial hydrolysates we conclude that the saccharide on C-3 is a β-D-glucoside, α-L-araboside, β-D-galactoside. 相似文献