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Salil K. Niyogi Thomas S. Soper Robert S. Foote Frank W. Larimer Richard J. Mural Sankar Mitra Eva H. Lee Richard Machanoff Fred C. Hartman 《Journal of biosciences》1987,11(1-4):203-214
Both Lys-166 and His-291 of ribulosebisphosphate carboxylase/oxygenase fromRhodospirillum rubrum have been implicated as the active-site residue that initiates catalysis. To decide between these two candidates, we resorted
to site-directed mutagenesis to replace Lys-166 and His-291 with several amino acids. All 7 of the position-166 mutants tested
are severely deficient in carboxylase activity, whereas the alanine and serine mutants at position 291 are ∼40% and ∼18% as
active as the native carboxylase, essentially ruling out His-291 in theRhodospirillum rubrum carboxylase (and by inference His-298 in the spinach enzyme) as a catalytically essential residue. The ability of some of
the mutant proteins to undergo carbamate formation or to bind either ribulosebisphosphate or a transition-state analogue remains
largely unimpaired. This implies that Lys-166 is not required for substrate binding; rather, the results corroborate the earlier
postulate that Lys-166 functions as an acid-base group in catalysis or in stabilizing a transition state in the reaction pathway. 相似文献
3.
R. Sankar P. S. Devamanoharan G. Raghupathi M. Krishnasamy C. S. Shyamala Devi 《Journal of biosciences》1987,12(3):267-271
Plumbagin was administered to rats at a concentration of 1,2,4,8 and 16 mg per kg body weight. After 24 h lipid peroxide levels
were found to decrease in subcellular fractions of liver. Plumbagin inhibited ascorbate and nicotinafde adenine dinucleotide
phosphate (reduced) dependent lipid peroxidation but was without any effect on cumene hydroperoxide dependent lipid peroxidation.
Injection of 16 mg of plumbagin per kg body weight was found to decrease liver total reduced glutathione and also fcrosomal
glucose-6-phosphatase. The results are discussed with reference to the anti- and prooxidant properties of plumbagin. 相似文献
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5.
Cloning of hydrogenase genes and fine structure analysis of an operon essential for H2 metabolism in Escherichia coli. 总被引:12,自引:10,他引:2 下载免费PDF全文
Escherichia coli has two unlinked genes that code for hydrogenase synthesis and activity. The DNA fragments containing the two genes (hydA and hydB) were cloned into a plasmid vector, pBR322. The plasmids containing the hyd genes (pSE-290 and pSE-111 carrying the hydA and hydB genes, respectively) were used to genetically map a total of 51 mutant strains with defects in hydrogenase activity. A total of 37 mutants carried a mutation in the hydB gene, whereas the remaining 14 hyd were hydA. This complementation analysis also established the presence of two new genes, so far unidentified, one coding for formate dehydrogenase-2 (fdv) and another producing an electron transport protein (fhl) coupling formate dehydrogenase-2 to hydrogenase. Three of the four genes, hydB, fhl, and fdv, may constitute a single operon, and all three genes are carried by a 5.6-kilobase-pair chromosomal DNA insert in plasmid pSE-128. Plasmids carrying a part of this 5.6-kilobase-pair DNA (pSE-130) or fragments derived from this DNA in different orientations (pSE-126 and pSE-129) inhibited the production of active formate hydrogenlyase. This inhibition occurred even in a prototrophic E. coli, strain K-10, but only during an early induction period. These results, based on complementation analysis with cloned DNA fragments, show that both hydA and hydB genes are essential for the production of active hydrogenase. For the expression of active formate hydrogenlyase, two other gene products, fhl and fdv are also needed. All four genes map between 58 and 59 min in the E. coli chromosome. 相似文献
6.
Antisera towards neurotensin (NT) and the structurally related peptide, LANT6, were used to characterize immunoreactive peptides and proteins in extracts of chicken tissues. A 17 kDa protein was identified by Western blotting as a potential precursor to NT and LANT6. However, the posttranslational processing of this common precursor appeared to be tissue specific, giving rise to disproportionate amounts of NT and LANT6, along with varying expression of a large molecular LANT6 (Mr, 15 kDa). The intestinal cells containing immunoreactive NT, LANT6, and large molecular LANT6 behaved similarly during fractionation by size and density. These activities also banded together in particles resembling vesicles during centrifugation of isotonic homogenates of tissue. These results suggest that chicken NT and LANT6 are biosynthesized as parts of the same precursor, the processing of which can give rise to a variety of products stored within secretory vesicles. 相似文献
7.
A new medium containing sodium tripolyphosphate, ethylenediaminetetraacetic acid disodium salt, dihydrate, sodium desoxycholate, supplemented with minerals and nutrient sources was developed, which was very effective in inhibiting swarming of Bacillus spp. This medium polyphosphate-EDTA-desoxycholate agar (PEDA) was compared with tryptone glucose extract agar (Oxoid) for determining the total plate count of various semi-conserved fishery products. While promoting discrete colony formation of Bacillus spp., PEDA provided good bacterial recovery. PEDA is recommended for isolation or enumeration of bacteria in food dominated by swarming Bacillus spp. 相似文献
8.
Nitesh Kumar Somlata Mohit Mazumder Priyanka Dutta Sankar Maiti Samudrala Gourinath 《PLoS pathogens》2014,10(9)
Entamoeba histolytica is a protist parasite that is the causative agent of amoebiasis, and is a highly motile organism. The motility is essential for its survival and pathogenesis, and a dynamic actin cytoskeleton is required for this process. EhCoactosin, an actin-binding protein of the ADF/cofilin family, participates in actin dynamics, and here we report our studies of this protein using both structural and functional approaches. The X-ray crystal structure of EhCoactosin resembles that of human coactosin-like protein, with major differences in the distribution of surface charges and the orientation of terminal regions. According to in vitro binding assays, full-length EhCoactosin binds both F- and G-actin. Instead of acting to depolymerize or severe F-actin, EhCoactosin directly stabilizes the polymer. When EhCoactosin was visualized in E. histolytica cells using either confocal imaging or total internal reflectance microscopy, it was found to colocalize with F-actin at phagocytic cups. Over-expression of this protein stabilized F-actin and inhibited the phagocytic process. EhCoactosin appears to be an unusual type of coactosin involved in E. histolytica actin dynamics. 相似文献
9.