全文获取类型
收费全文 | 4890篇 |
免费 | 207篇 |
国内免费 | 193篇 |
出版年
2023年 | 40篇 |
2022年 | 65篇 |
2021年 | 105篇 |
2020年 | 104篇 |
2019年 | 121篇 |
2018年 | 108篇 |
2017年 | 108篇 |
2016年 | 80篇 |
2015年 | 98篇 |
2014年 | 142篇 |
2013年 | 280篇 |
2012年 | 113篇 |
2011年 | 198篇 |
2010年 | 135篇 |
2009年 | 226篇 |
2008年 | 261篇 |
2007年 | 220篇 |
2006年 | 217篇 |
2005年 | 197篇 |
2004年 | 188篇 |
2003年 | 195篇 |
2002年 | 154篇 |
2001年 | 107篇 |
2000年 | 121篇 |
1999年 | 107篇 |
1998年 | 103篇 |
1997年 | 104篇 |
1996年 | 109篇 |
1995年 | 106篇 |
1994年 | 101篇 |
1993年 | 100篇 |
1992年 | 81篇 |
1991年 | 71篇 |
1990年 | 86篇 |
1989年 | 55篇 |
1988年 | 64篇 |
1987年 | 52篇 |
1986年 | 54篇 |
1985年 | 67篇 |
1984年 | 80篇 |
1983年 | 42篇 |
1982年 | 57篇 |
1981年 | 57篇 |
1980年 | 43篇 |
1979年 | 44篇 |
1978年 | 31篇 |
1977年 | 21篇 |
1976年 | 21篇 |
1975年 | 10篇 |
1973年 | 10篇 |
排序方式: 共有5290条查询结果,搜索用时 15 毫秒
1.
Wendy A. Douglass Robert H. Hyland Christopher D. Buckley Aymen Al-Shamkhani Jacqueline M. Shaw Sarah L. Scarth David L. Simmons S.K.Alex Law 《FEBS letters》1998,440(3):125
The cysteine-rich region (CRR) of the β2 integrin subunit was replaced by that of β1 to give the chimera β2NV1. β2NV1 can combine with αL to form a variant leukocyte-function-associated antigen (LFA)-1 on COS cell surface, suggesting that the specificity of the β2 interaction with αL does not lie in the CRR. Unlike those expressing wild-type LFA-1, COS cells expressing αLβ2NV1 are constitutively active in intercellular adhesion molecule (ICAM)-1 adhesion. These results suggest that activation of LFA-1 involves the release of an intramolecular constraint, which is maintained, in part, by the authentic β2 CRR. 相似文献
2.
Steven J. Tucker David McClelland Kristina Sep?i? Roderick H. Scott 《生物化学与生物物理学报:生物膜》2003,1614(2):171-181
The ability of two alkyl pyridinium sponge toxin preparations (poly-APS and halitoxin) to form transient pores/lesions in cell membranes and allow transfection of plasmid cDNA have been investigated using HEK 293 cells. Poly-APS and halitoxin preparations caused a collapse in membrane potential, reductions in input resistance and increased Ca2+ permeability. At least partial recovery was observed after poly-APS application but recovery was more rarely seen with halitoxin. The transfection with plasmid cDNAs for an enhanced green fluorescent protein (EGFP) and human tumour necrosis factor receptor 2 (TNFR2) was assessed for both toxin preparations and compared with lipofectamine. Stable transfection was achieved with poly-APS although it was less efficient than lipofectamine. These results show that viable cells transfected with alien cDNA can be obtained using novel transient pore-forming alkyl pyridinium sponge toxins and a simple pre-incubation protocol. This provides the first proof of principle that pore-forming alkyl pyridinium compounds can be used to deliver cDNA to the intracellular environment without permanently compromising the plasma membrane. 相似文献
3.
Ice crystal formation temperature was determined in the region of the crown in one group of 7-day-old intact unhardened high-salt plants of winter wheat (Triticum aestivum L. cv. Weibulls Starke II) with TA (Thermal Analysis) and DTA (Differential Thermal Analysis) methods. After exposure of another group of plants, grown for the first 7 days in the same way as the first group, to various sub-zero temperatures (-1 to 5°C), influx in roots of Rb+(86Rb+) and Ca2+(45Ca2+) and contents of K+ and Ca2+ were determined at intervals during 7 days of recovery. Ice crystal formation in the crown tissue was probably extracellular and took place at about -4°C. There was a large loss of K+ from the roots after treatment at sub-zero temperatures. This loss increased as the temperature of the sub-zero treatment decreased. During recovery, roots of plants exposed to -1, -2 and -3°C gradually reabsorbed K+. Reabsorption of K+ in roots of plants exposed to -4°C was greatly impaired. Rb+ influx decreased and Ca2+ influx increased after sub-zero temperature treatments of the plants. Active Rb+ influx mechanisms and active extrusion of Ca2+ were impaired or irreversibly damaged by the exposure. While Rb+ influx mechanisms were apparently repaired during recovery in plants exposed to temperatures down to -3°C, Ca2+ extrusion mechanisms were not. The temperature for ice crystal formation in the region of the crown tissue coincides with the temperature at which the plants lost the ability to reabsorb K+ and to repair Rb+ influx mechanisms during the recovery period. Plants were lethally damaged at temperatures below ?4°C. 相似文献
4.
A strain of Methylomonas albus BG8WM, a type 1 obligate methanotroph, which had been maintained for 2 ycars by serial passage on solid medium containing methanol as a carbon source was found to mutate at a frequency of 10-5-10-6 to resistance to dichloromethane (DCMR), the parental strain BG8 did not give rise to DCMR colonies. DCMR strains were no longer capable of growth on methane as a carbon cource and exhibited greatly reduced or undetectable methane mono-oxygenase activity. The mutants fell into three groups on the basis of SDS-PAGE analysis of the polypeptide profiles of the particulate fraction of cell extracts. One or more of four polypeptides of Mr 70,000, 50,000, 25,000 and 23,000 were implicated as being components of the methane mono-oxygenase. Spontaneous reversion to growth on methane and sensitivity to dichloromethane occurred at a frequency of about 10-8. The loss of methane mono-oxygenase activity was not associated with loss of the resident 55 kb plasmid.Abbreviations DCMR
dichloromethane-resistant
- SDS-PAGE
sodium dodecyl sulphate polyacrylamide gel electrophoresis
- NMS
nitrate minimal salts medium 相似文献
5.
The genes encoding the six polypeptide components of the alkene monooxygenase from Xanthobacter Py2 have been sequenced. The predicted amino acid sequence of the first ORF shows homology with the iron binding subunits of binuclear non-haem iron containing monooxygenases including benzene monooxygenase, toluene 4-monooxygenase (>60% sequence similarity) and methane monooxygenase (>40% sequence similarity) and that the necessary sequence motifs associated with iron co-ordination are also present. Secondary structure prediction based on the amino acid sequence showed that the predominantly α-helical structure that surrounds the binuclear iron binding site was conserved allowing the sequence to be modelled on the co-ordinates of the methane monooxygenase α-subunit. Significant differences in the residues forming the hydrophobic cavity which forms the substrate binding site are discussed with reference to the differences in reaction specificity and stereospecificity of binuclear non-haem iron monooxygenases. 相似文献
6.
Y. Jiang 《Biochimica et Biophysica Acta (BBA)/General Subjects》1994,1201(1):76-84
Proteolysis of the hydroxylase component of soluble methane monooxygenase (MMO) with trypsin yielded a protein which retained 50% activity in a standard MMO assay. In an H2O2-driven assay, in which H2O2 replaced two of the protein components, NADH and O2 used in the standard assay, the proteolysed hydroxylase retained full activity for ethane, propane and propene, but had a 2–3 fold increase with methane as substrate. Several crosslinking reagents have been tested for their ability to stabilise the proteolysed form of the hydroxylase. Using polyoxyethylene bis(imidazolyl carbonyl) (Mr 3350) as the crosslinking agent, increased thermostability of the hydroxylase was observed. Activated methoxypolyethylene glycol (Mr 5000) was used to modify the hydroxylase which was now soluble in organic solvents as well as water and could be activated by H2O2. The glycol-modified hydroxylase functioned well in organic solvents in the catalysis of propene oxidation. 相似文献
7.
Structure and Functions of Channel-Forming Peptides: Magainins,Cecropins, Melittin and Alamethicin 总被引:26,自引:0,他引:26
B. Bechinger 《The Journal of membrane biology》1997,156(3):197-211
8.
Abstract Both the intracellular and the extracellular concentration of cyclic AMP increases logarithmically in synchronously growing cultures of Escherichia coli . Thus, cyclic AMP by itself cannot regulate growth and division of the bacterium during the cell cycle. 相似文献
9.
Antioxidant effect of diethyldithiocarbamate on microsomal lipid peroxidation assessed by low-level chemiluminescence and alkane production 总被引:3,自引:0,他引:3
Different thiol-containing compounds, such as diethyldithiocarbamate (DDC), glutathione, penicillamine, and dithioerythritol have been chosen to study their effect on ascorbate/Fe-ADP-induced lipid peroxidation, detected by low-level chemiluminescence and alkane production. In the concentration range used, these thiols exerted a temporary protection against lipid peroxidation by lengthening the induction period; after overcoming this induction period, no substantial inhibition of either chemiluminescence or alkane production was observed. DDC was effective in protecting against lipid peroxidation in the nanomolar range, whereas the group of other thiol-containing molecules operated in the millimolar range. 相似文献
10.
Mamta Singhvi Dipti Joshi Shalaka Gaikaiwari Digambar V. Gokhale 《Indian journal of microbiology》2010,50(1):97-100
Method for production and regeneration of Lactobacillus delbrueckii protoplasts are described. The protoplasts were obtained by treatment with a mixture of lysozyme and mutanolysin in protoplast
buffer at pH 6.5 with different osmotic stabilizers. The protoplasts were regenerated on deMan, Rogosa and Sharpe (MRS) with
various osmotic stabilizers. Maximum protoplast formation was obtained in protoplast buffer with sucrose as an osmotic stabilizer
using a combination of lysozyme (1 mg/ml) and mutanolysin (10 μg/ml). Maximum protoplast regeneration was obtained on MRS
medium with sucrose (0.5 M) as an osmotic stabilizer. The regeneration medium was also applicable to other species of lactobacilli
as well. This is, to our knowledge, the first report on protoplast formation and efficient regeneration in case of L. delbrueckii. 相似文献