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61.
Genetic transformation of flax (Linum usitatissimum) by Agrobacterium tumefaciens: regeneration of transformed shoots via a callus phase 总被引:1,自引:0,他引:1
Genetic transformation of flax (Linum usitatissimum) has been achieved using an A. tumefaciens strain carrying a non-oncogenic Ti plasmid-derived vector containing a chimaeric npt-II gene and a wild type nopaline synthase gene. Fertile, transformed shoots were most easily obtained from Kmr callus developing on hypocotyl sections. The totipotency of the Kmr callus was dependent upon its origin. T-DNA was visualised by Southern blotting in all Kmr tissues. Efficient expression of nopaline synthase and the chimaeric npt-II gene was found in transformed Kmr callus and regenerated shoots.Abbreviations
npt-II
neomycin phosphotransferase II gene
- NPT-II
neomycin phosphotransferase II
- nos
nopaline synthase gene promoter
- Kmr
kanamycin resistant
- BAP
6-benzylaminopurine
- NAA
-naphthaleneacetic acid
- MSD4×2
medium D4×2 based on Murashige & Skoog medium (see Scott & Draper, 1987) 相似文献
62.
63.
The fluorescence of the lipophilic prbe N-phenyl-1-naphthylamine (NPN) bound to intact cells of Escherichia coli is quenched by the addition of glucose, succinate,
-lactate, pyruvate, formate and glycerol. Partial recovery of fluorescence occurs on anaerobiosis. Use of mutants with defects in the ATP synthase or the respiratory chain show that quenching of fluorescence may be energized either by ATP hydrolysis or by substrate oxidation through the respiratory chain. Permeabilization of the outer membrane by treatment of intact cells with EDTA, or use of a mutant with an outer membrane permeable to lipophilic substances, results in a more rapid binding of NPN and in a decrease in quenching observed on substrate addition. NPN binds rapidly to everted membrane vesicles, but does not respond to membrane energization. It is proposed that inner membrane energization in intact cells alters the binding or environment of NPN in the outer membrane. The fluorescence recovery which occurs on anaerobiosis has two components. One component represents a reversal of the changes which occur on membrane energization. The other component of the fluorescence change is insensitive to the uncoupler CCCP and resembles the behaviour of NPN with everted membrane vesicles. It is suggested that a portion of the fluorescence events seen with NPN involves a response of the probe to changes in the inner membrane. 相似文献
64.
S Rogne G Skretting F Larsen O Myklebost B Mev?g L A Carlson L Holmquist E Gjone H Prydz 《Biochemical and biophysical research communications》1987,148(1):161-169
We have isolated cDNA clones coding for human lecithin:cholesterol acyl transferase (LCAT) from a liver-specific cDNA library by the use of two oligonucleotide probes based on the protein sequence. The clones span the sequence coding for the entire secreted LCAT, the 3' untranslated sequence and 12 amino acids of the signal peptide. The peptide sequence contains the conserved active site of serine lipases within a hydrophobic domain, flanked by a possible amphipatic alpha-helix. Only one gene for LCAT could be detected in genomic blots. We have used the cDNA as a probe to analyse the LCAT gene in patients suffering from LCAT deficiency and fish eye disease. No rearrangements or abnormal gene fragments were detected in these patients. 相似文献
65.
66.
A Kryski T S Larsen I Ramírez D L Severson 《Canadian journal of physiology and pharmacology》1987,65(2):226-229
Triacylglycerol lipase activity with a pH optimum of 5 was present in homogenates of myocardial cells from rat heart. Acid lipase activity was inhibited by serum, heparin, and increased ionic strength. Methylamine, a lysosomotropic agent, did not inhibit the basal or isoproterenol-stimulated rate of endogenous lipolysis as measured by glycerol output from control myocytes. Similarly, accelerated rates of glycerol output that are a consequence of an elevation in the intracellular stores of triacylglycerols in myocytes from diabetic rat hearts and from myocytes prepared with free fatty acids in the isolation solutions were not reduced by methylamine. Therefore, the acid lysosomal triacylglycerol lipase must not be involved in the mobilization of endogenous triacylglycerols in myocardial cells from rat heart. 相似文献
67.
Osmoregulation in Escherichia coli by accumulation of organic osmolytes: betaines,glutamic acid,and trehalose 总被引:42,自引:0,他引:42
Like other cyanobacteria, Chlorogloeopsis fritschii contained as major lipid classes monogalactosyldiacyl-glycerols, digalactosyldiacylglycerols, sulfoquinovosyldiacylglycerols and diacylglycerophosphoglycerols. In addition to these lipid classes this cyanobacterium also contained small amounts of diacylglycerophosphocholines and sterols, predominantly lanosterol, thus showing similarity to photosynthetic eukaryotes. Dark incubated cells contained larger proportions of the latter two lipid classes than light grown cells. Like other prokaryotes, C. fritschii lacked linolenic acid (18:3) in its lipids. Lipids from the thylakoids were richer in palmitoleic acid (16:1) than those of whole cells. There was no effect of light on the patterns of constituent fatty acids of lipids from C. fritschii, in contrast to photosynthetic eukaryotes.Abbreviations MGDG
Monogalactosyldiacylglycerols
- PA
Phosphatidic acids
- PE
Diacylglycerophosphoethanolamines
- PG
Diacylglycerophosphoethanolamines
- DGDG
Digalactosyldiacylglycerols
- SQDG
Sulfoquinovosyldiacylglycerols
- PC
Diacylglycerophosphocholines 相似文献
68.
TenVibrio anguillarum strains were selected for an immunoelectrophoretic study. Evidence was provided for existence of two new K antigens which displayed cross-reactivity. The importance of an exact characterization of surface antigens inV. anguillarum is considered. 相似文献
69.
Rates of mutation to growth factor autonomy and tumorigenicity differ in hematopoietic stem and precursor cells expressing the multilineage colony-stimulating factor gene. 总被引:2,自引:0,他引:2
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C Laker N Kluge C Stocking U Just M J Franz W Ostertag J F DeLamarter M Dexter E Spooncer 《Molecular and cellular biology》1989,9(12):5746-5749
At least two separate but interdependent events are required to attain autonomous growth as a consequence of ectopic expression of the multilineage colony-stimulating factor gene in hematopoietic progenitor cells. The rate at which the second event occurs is more than 3 orders of magnitude higher in precursor cell lines (FDC-P1 or FDC-P2) than in stem cell lines (FDC-Pmix). Autonomous, but not density-dependent, growth is tightly coupled to tumorigenicity in precursor cells; however, neither growth-factor-independent nor autonomously growing stem cell lines are tumorigenic. 相似文献
70.
Mary Welch Baillargeon Raymond G. Bistline Jr. Philip E. Sonnet 《Applied microbiology and biotechnology》1989,30(1):92-96
Summary Three strains of Geotrichum candidum (ATCC 34614, NRRL Y-552 and NRRL Y-553) were examined for lipase production and activity. Variables including medium, pH, temperature, agitation rate and incubation time were examined to define the optimal culture conditions. Growth on oil in complex medium at 30°C, 300 rpm, and pH 7 produced maximal lipase activity. Fatty acid specificity of these strains and of two crude G. candidum enzyme preparations (lipase 26557 RP, Rhône Poulenc and lipase GC-4, Amano) was measured using equimolar mixtures of methyl or butyl esters of palmitic and oleic acids. The lipase from NRRL Y-553 and lipase 26557 RP displayed preferential specificity for hydrolyzing oleic acid esters, while the lipases from ATCC 34614, NRRL Y-552 and lipase GC-4 failed to discriminate between plamitic and oleic acids. 相似文献