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1.
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
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Controlled partial restriction digestions of DNA by competition with modification methyltransferases 总被引:3,自引:0,他引:3
Competitive reactions, using defined ratios of DNA restriction methyltransferase to endonuclease, are shown to result in reliable partial restriction digests of DNA. This method is suitable over a wide range of DNA concentrations and works on DNA in liquid or embedded in agarose. Simultaneous methylase/endonuclease reactions using endonucleases that cleave human DNA very infrequently, such as ClaI or NotI, should generate very large discrete partial DNA fragments suitable for physical mapping in the million base-pair range. Another possible application of methylase/endonuclease competitive reactions is the production of defined partial digests for making cosmid, lambda, or other genomic libraries. 相似文献
5.
D B McClelland 《BMJ (Clinical research ed.)》1990,300(6716):35-37
6.
Cecilia PC Soh Alastair SR Donald James Feeney Walter TJ Morgan Winifred M Watkins 《Glycoconjugate journal》1989,6(3):319-332
The tetrasaccharides GalNAcß1-4[NeuAc2-3]Galß1-4Glc and GalNAcß1-4[NeuAc2-3]Galß1-4GlcNAc were synthesised by enzymic transfer of GalNAc from UDP-GalNAc to 3-sialyllactose (NeuAc2-3Galß1-4Glc) and 3-sialyl-N-acetyllactosamine (NeuAc2-3Galß1-4GlcNAc). The structures of the products were established by methylation and1H-500 MHz NMR spectroscopy. In Sda serological tests the product formed with 3-sialyl-N-acetyllactosamine was highly active whereas that formed with 3-sialyllactose had only weak activity. 相似文献
7.
M. A. L. Smith L. Art Spomer M. J. Meyer M. T. McClelland 《Plant Cell, Tissue and Organ Culture》1989,19(2):91-102
Microcomputerized video image analysis was adapted for rapid, objective, and non-intrusive quantification of shoot growth and development for plants growing in vitro. Custom-developed staging arrangements were essential to insure accurate viewing and representation of the plants in each of three standard culture vessels. Shoot length measurements from digitized culture images were strongly correlated with length measured manually ex vitro. Image analysis weighted density measurements of proliferating microcultures (even with irregular growth habits) provided a reliable indicator of shoot culture fresh weight. Non-destructive time course evaluations of growth rate and quality were demonstrated.Abbreviations FW
fresh weight
- WD
weighted density 相似文献
8.
Anna L. Ballard Alastair G. McEwan David J. Richardson J. Baz Jackson Stuart J. Ferguson 《Archives of microbiology》1990,154(3):301-303
Rhodobacter capsulatus strain BK5 possesses a membrane bound respiratory nitrate reductase rather than the periplasmic enzyme found in other strains. The enzyme in strain BK5 is shown to be both functionally and structurally related to the nitrate reductase of Paracoccus denitrificans and Escherichia coli.Abbreviation TMAO
trimethylamine-N-oxide 相似文献
9.
Bruno W. S. Sobral Rhonda J. Honeycutt Alan G. Atherly Michael McClelland 《Plant Molecular Biology Reporter》1990,8(4):253-275
TheOryza sativa (rice) genome is small (600 to 900 megabase pairs) when compared to that of other monocotyledonous plants. Rice was the first
of the major cereals to be successfully transformed and regenerated. An RFLP map with approximately 300 markers is readily
available, and the DNA content per map unit is only two to three times that ofArabidopsis thaliana. Rice is also the main staple food for the majority of peoples in the world. We developed techniques for the preparation
of intact genomic DNA from Indica and Japonica subspecies of rice, used statistical methods to determine which restriction
endonucleases are rare-cutting, and used pulsed-field gel electrophoresis (PFE) to separate large fragments of rice DNA. Southern
hybridization to blotted rice PFE gels was used to show that the digests were complete. The long-term goal of our work is
to generate an integrated genetic/physical map for the rice genome, as well as helping to establish rice as a model for map-based
gene cloning and genome analysis. 相似文献
10.
Summary The effects of a non-ionic surfactant, Pluronic F-68, on growth of microbial cell cultures have been studied. Growth ofSaccharomyces cerevisiae at 30°C or 37°C as measured by viable cell counts was unaffected by culture with pluronic. However, corresponding absorbance measurements forS. cerevisiae incubated with 5–10% pluronic were lower than controls at both temperatures. Absorbance ofE. coli cultures was also significantly reduced by incubation with 5.0–10.0% pluronic at both temperatures although viable counts again revealed no significant inhibition of growth. 相似文献