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61.
Robin E. Everts Serge A. Versteeg Corinne Renier Francoise Vignaux Peter C. Groot Jan Rothuizen Bernard A. van Oost 《Mammalian genome》2000,11(9):741-747
Genomic Representational Difference Analysis (gRDA) is a subtractive DNA method to clone the differences between two related
genomes, called tester and driver. We have evaluated this method to obtain polymorphic DNA markers for pedigree dogs. Amplified
size-selected genomic restriction fragments (amplicons) of two dog littermates were repeatedly hybridized to each other in
order to remove (subtract) those restriction fragments common to both sibs. Already after two rounds of subtractive hybridization,
a clear enrichment of presumably tester-specific restriction fragments was observed, which was even more pronounced after
the third round of subtraction. A plasmid library of 3000 recombinant clones was constructed of the second round and of the
third round difference product. DNA sequence determination of randomly chosen clones of each difference product showed that
approximately 1000 unique clones were obtained in the second-round difference product and approximately 500 in the third-round
difference product. About half of the clones identified in the second-round difference product were also present in the third-round
difference product. Of the second-round difference product, 39 different gRDA fragments could be identified, of which 21 were
tester specific. In the third-round difference product, 22 different gRDA fragments were identified, of which 18 were tester
specific. There were 13 fragments in common, resulting in a total of 48 different fragments. In order to establish the localization
of these markers, we performed mapping using the dog radiation hybrid panel RHDF5000. Of 39 mapped clones, 29 were mapped
to 20 existing RH groups, and 10 remained unlinked. It is concluded that gRDA is suitable to generate DNA markers to track
disease genes within lines of pedigree dogs.
Received: 26 April 2000 / Accepted: 11 May 2000 相似文献
62.
Flavans and procyanidins from the seeds of different grape varieties were separated and identified using HPLC techniques. The compounds identified were (+)-catechin and (?)-epicatechin, dimeric procyanidins B1, B2, B3 and B4, trimeric procyanidin C2 and gallic acid. During maturation of the grape berries, the flavan-3-ol content fell in the seeds whereas procyanidin levels increased. This suggests an interrelationship between the compounds. There was also evidence of varietal differences in the amounts of phenolic compounds in grape seeds. 相似文献
63.
Endurance training affects myosin heavy chain phenotype in regenerating fast-twitch muscle 总被引:2,自引:0,他引:2
Bigard Xavier A.; Janmot Chantal; Merino Daniele; Lienhard Francoise; Guezennec Yannick C.; D'Albis Anne 《Journal of applied physiology》1996,81(6):2658-2665
Bigard, Xavier A., Chantal Janmot, Danièle Merino,Françoise Lienhard, Yannick C. Guezennec, and Anne D'Albis.Endurance training affects myosin heavy chain phenotype inregenerating fast-twitch muscle. J. Appl.Physiol. 81(6): 2658-2665, 1996.The aim of thisstudy was to analyze the effects of treadmill training (2 h/day, 5 days/wk, 30 m/min, 7% grade for 5 wk) on the expression of myosinheavy chain (MHC) isoforms during and after regeneration of afast-twitch white muscle [extensor digitorum longus (EDL)]. Male Wistar rats were randomly assigned to a sedentary(n = 10) or an endurance-trained (ET;n = 10) group. EDL muscle degeneration and regeneration were induced by two subcutaneous injections of a snaketoxin. Five days after induction of muscle injury, animals were trainedover a 5-wk period. It was verified that ~40 days after venomtreatment, central nuclei were present in the treated EDL muscles fromsedentary and ET rats. The changes in the expression of MHCs in EDLmuscles were detected by using a combination of biochemical andimmunocytochemical approaches. Compared with contralateral nondegenerated muscles, relative concentrations of types I, IIa, andIIx MHC isoforms in ET rats were greater in regenerated EDL muscles(146%, P < 0.05; 76%,P < 0.01; 87%,P < 0.01, respectively). Their elevation corresponded to a decreasein the relative concentration of type IIb MHC (36%,P < 0.01). Although type I accountedfor only 3.2% of total myosin in regenerated muscles from the ETgroup, the cytochemical analysis showed that the proportion of positive staining with the slow MHC antibody was markedly greater in regenerated muscles than in contralateral ones. Collectively, these results demonstrate that the regenerated EDL muscle is sensitive to endurance training and suggest that the training-induced shift in MHC isoforms observed in these muscles resulted from an additive effect of regeneration and repeated exercise. 相似文献
64.
Francoise H Routier Michael J Davies Klaus Bergemann Elizabeth F Hounsell 《Glycoconjugate journal》1997,14(2):201-207
A humanized IgG antibody (D1.3) which retains murine complementarity determining regions specific for the antigen lysozyme
has been expressed in CHO-DUKX cells. Heavy and light chain containing plasmids were co-transfected into CHO-DUKX cells and
stable clones were grown in DMEM/F12 medium supplemented with 5% foetal calf serum. D1.3 antibody was purified from culture
supernatants by Protein G chromatography. With the recombinant D1.3 antibody as a model, this cell culture system was shown
to glycosylate the IgG Fc region in a similar manner to IgG isolated from serum. The neutral, core fucosylated biantennary
oligosaccharides found are present in serum IgG and no novel carbohydrate sequences were detected. The degree of terminal
agalactosylation was also similar to normal serum, in contrast to the increased levels found in rheumatoid serum. Furthermore,
those oligosaccharides which lack only one terminal Gal are exclusively galactosylated on the GlcNAc(β1,2) Man(α1,6) Man(β1,4) antenna. Unambiguous identification of the exact glycosylation pattern of the antibody was carried out by a combination
of specific exoglycosidase digestions, gel permeation chromatography of 2-aminobenzamide derivatives, high pH anion exchange
chromatography and methylation analysis followed by gas–liquid chromatography-mass spectrometry. Abbreviations: CDR, complementarity
determining region; CHO, chinese hamster ovary; GPC, gel permeation chromatography; 2-AB, 2-aminobenzamide; HPAEC-PAD, high
pH anion exchange chromatography with pulsed amperometric detection; GC-MS, gas chromatography with mass spectrometry analysis;
PNGase F, peptide-N-glycosidase F; PGC, porous graphitized carbon column; RAAM, reagent array analysis method; NeuAc: N-acetylneuraminic
acid; NeuGc: N-glycolylneuraminic acid
This revised version was published online in November 2006 with corrections to the Cover Date. 相似文献
65.
Fathallah Hassana; Sauvage Monique; Romero Jose R.; Canessa Mitzy; Giraud Francoise 《American journal of physiology. Cell physiology》1997,273(4):C1206
We have previously shown that a pretreatment with phorbol12-myristate 13-acetate (PMA), an activator of protein kinase C (PKC),reduced deoxygenation-induced K+loss and Ca2+ uptake and preventedcell dehydration in sickle anemia red blood cells (SS cells) (H. Fathallah, E. Coezy, R.-S. De Neef, M.-D. Hardy-Dessources, and F. Giraud. Blood 86: 1999-2007,1995). The present study explores the detailed mechanism of thisPMA-induced inhibition. The main findings are, first, the detection ofPKC and PKC in normal red blood cells and the demonstration that both isoforms are expressed at higher levels in SS cells. The -isoform only is translocated to the membrane and activated by PMAand by elevation of cytosolicCa2+. Second, PMA is demonstratedto activate Ca2+ efflux indeoxygenated SS cells by a direct stimulation of the Ca2+ pump. PMA, moreover, inhibitsdeoxygenation-induced, charybdotoxin-sensitive K+ efflux in SS cells. Thisinhibition is partly indirect and explained by the reduceddeoxygenation-induced rise in cytosolicCa2+ resulting fromCa2+ pump stimulation. However, asignificant inhibition of theCa2+-activatedK+ channels(KCa channels) by PMA can also bedemonstrated when the channels are activated byCa2+ plus ionophore, underconditions in which the Ca2+ pumpis operating near its maximal extrusion rate, but swamped byCa2+ plus ionophore. The data thussuggest a PKC-mediated phosphorylation both of theCa2+ pump and of theKCa channel or an auxiliaryprotein. 相似文献
66.
67.
Temperature-Sensitive Initiation of Chromosome Replication in a Mutant of Bacillus subtilis 总被引:14,自引:7,他引:7
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A mutant of Bacillus subtilis Ts37 has been isolated in which deoxyribonucleic acid (DNA) synthesis is inhibited at high temperature. The results presented here indicate that the process of initiation of DNA replication is temperature sensitive in this mutant. After shifting to 45 C, DNA increases 40 to 50% before synthesis ceases; an inhibition of protein synthesis permits an equivalent amount of DNA to be synthesized. A density shift experiment coupled with a marker frequency analysis shows that DNA synthesized at 45 C is highly enriched in the markers situated at the end of the chromosome. Transforming DNA extracted from a culture which has been incubated at 45 C exhibits the relative transforming efficiency for origin and terminus markers characteristic of completed chromosomes. After a shift back from 45 C to 30 C, reinitiation appears to occur always in the same region of the bacterial chromosome; in addition, replication as well as cell division is synchronized. 相似文献
68.
Heat-induced changes in respiratory pathways and mitochondrial structure during microcycle conidiation of Neurospora crassa 总被引:1,自引:0,他引:1
Mehrbanou Michéa-Hamzehpour Francoise Grange The-Can Ton That Gilbert Turian 《Archives of microbiology》1980,125(1-2):53-58
Changes in both respiratory pathways and mitochondrial structure of Neurospora crassa occurred under conditions of microcycle conidiation. Upon heat-treatment at 46°C, conidia developed a highly cyanide-insensitive, hydroxamate-sensitive respiration associated with morphological alterations in mitochondrial membranes; such changes were time-dependent. When heat-treated conidia were shifted down to 25°C, the alternate, hydroxamate-sensitive respiration decreased significantly, paralleling the recovery of well-cristated mitochondria with an electron-dense matrix in the germ tubes. The decrease in hydroxamate-sensitivity was associated with two periods of increase in cyanide sensitivity corresponding to the events of germination and precocious proconidial budding. 相似文献
69.
70.