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1.
2.
Electron microscopy of single-stranded structures in the DNA of competent Haemophilus influenzae cells. 总被引:4,自引:3,他引:1
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Chromosomal DNAs from exponential-phase and competent cells of Haemophilus influenzae were examined by electron microscopy to determine whether the chromosome undergoes structural changes during competence development. Single-stranded gaps and single-stranded tails formed in chromosomal DNA during competence development. The generation of gaps was dependent on the rec-2 function. Since the rec-2 mutant is defective in the translocation of donor DNA, it was inferred that the gaps were involved in the translocation step of transformation. The generation of single-stranded tails was independent of the rec-1 and rec-2 genes. Therefore, these structures were assumed to play no direct role in the interaction of donor and recipient DNAs during transformation. Gaps were preferentially associated with a readily denaturable, possibly A + T-rich fraction of the genome. This finding raised the possibility that hot spots for transformation might be associated with A + T-rich DNA. 相似文献
3.
Previous studies suggested that rabbit liver microsomes contain cytochrome P-450 monooxygenase(s) with low affinity for (omega-1)-hydroxylation and high affinity for omega-hydroxylation of prostaglandins (Theoharides, A. D., and Kupfer, D. (1981) J. Biol. Chem. 256, 2168-2175). The current investigation describes the isolation from livers of untreated rabbits of a cytochrome P-450 catalyzing, with regioselectivity, the omega-hydroxylation of prostaglandins E1 and E2. The isolation of the enzyme involved enrichment of the omega-hydroxylase activity by polyethylene glycol 8000 fractionation, followed by ion-exchange high performance liquid chromatography. Based on Mr of 59,000-60,000 from sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the isolated enzyme is referred to as P-450 form 7. This P-450 exhibits a low spin spectrum (lambda max = 417 nm) and a difference spectrum of the CO-reduced complex versus reduced (lambda max = 451 nm). For catalytic activity, the P-450 form 7 was reconstituted with NADPH-P-450 reductase, cytochrome b5, and lipid. There was no activity in the absence of the reductase, and deletion of cytochrome b5 yielded a minimal amount of product (heme could not substitute for cytochrome b5), demonstrating an absolute requirement for these components. 相似文献
4.
The effects of methylcholanthrene (MC) treatment of male rats on the regioselectivity of hydroxylation of prostaglandins E1 and E2 (PGE1 and PGE2) by liver microsomes, supplemented with NADPH or H2O2, was examined. In the presence of NADPH, control microsomes catalyzed the hydroxylation at omega-1 (C19) and at omega-(C20) sites with minimal formation of novel monohydroxy metabolites of PGE1 and PGE2, referred to as compounds X1 and X2, respectively. Similarly, H2O2 supported the 19-hydroxylation and the formation of compounds X1 and X2, but yielded only minimal amounts of 20-hydroxy products. With NADPH, MC-treated microsomal incubations demonstrated only minor quantitative change in the 19- and 20-hydroxylation as compared with controls, but showed a 7- to 11-fold increase in formation of compound X1 and a 10-fold increase in formation of X2. By contrast with H2O2, MC-treatment increased by about 3-fold the 19- and 20-hydroxylation of PGE1 and by 35- to 46-fold the formation of X1; similarly, there was an approximate 2-fold increase in 19- and 20-hydroxylation of PGE2 and a 10-fold increase in formation of X2. These findings suggest that several monooxygenases are involved in catalyzing the hydroxylation at the various sites of the PGE molecule. Inhibitors of monooxygenases (SKF 525A, alpha-naphthoflavone, and imidazole derivatives) provided further evidence that the hydroxylation at the three sites of PGEs is catalyzed by different P-450 monooxygenases. It is striking that the inhibitors had a much lesser effect on the 20-hydroxylation of PGE1 as compared with other sites of hydroxylation. Structural identification of compounds X1 and X2 was elucidated as follows. Resistance of the PGB derivative of X1 to periodate oxidation and mass fragmentation analysis of the t-butyldimethylsilyl ether methyl ester, placed the hydroxylation at C17 or C18. Finally, mass fragmentation of trimethylsilyl ether methyl ester PGB derivatives of X1 and X2 provided conclusive evidence that X1 and X2 are 18-hydroxy-PGE1 and 18-hydroxy-PGE2, respectively. The above findings indicate that the high regioselectivity of hydroxylation of PGE1 and PGE2, resulting in the formation of 18-hydroxy-PGE1 and 18-hydroxy-PGE2, respectively, is catalyzed by P-450 isozyme(s) which are induced by MC, possibly by P-450c. 相似文献
5.
mtDNA diversity in rhesus monkeys reveals overestimates of divergence time and paraphyly with neighboring species 总被引:4,自引:0,他引:4
Reconstructions of the human-African great ape phylogeny by using
mitochondrial DNA (mtDNA) have been subject to considerable debate. One
confounding factor may be the lack of data on intraspecific variation. To
test this hypothesis, we examined the effect of intraspecific mtDNA
diversity on the phylogenetic reconstruction of another Plio- Pleistocene
radiation of higher primates, the fascicularis group of macaque (Macaca)
monkey species. Fifteen endonucleases were used to identify 10 haplotypes
of 40-47 restriction sites in M. mulatta, which were compared with similar
data for the other members of this species group. Interpopulational,
intraspecific mtDNA diversity was large (0.5%- 4.5%), and estimates of
divergence time and branching order incorporating this variation were
substantially different from those based on single representatives of each
species. We conclude that intraspecific mtDNA diversity is substantial in
at least some primate species. Consequently, without prior information on
the extent of genetic diversity within a particular species, intraspecific
variation must be assessed and accounted for when reconstructing primate
phylogenies. Further, we question the reliability of hominoid mtDNA
phylogenies, based as they are on one or a few representatives of each
species, in an already depauperate superfamily of primates.
相似文献
6.
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8.
Pacific giant salamanders (Dicamptodon) rank among the largest terrestrial caudates. Their ontogeny produces two distinct morphs—larval‐neotenic and metamorphosed—which differ in many morphological traits. We identified changes that are initiated by metamorphosis (distinguishing transformed from neotenic specimens) and also recognized age‐related changes occurring irrespective of transformation. During metamorphosis, specimens remodel the palate, rearrange the vomerine dentition, expand the maxilla, broaden the cheek, foreshorten the posterior skull table and develop specific serrated suture patterns in the dermal bones. Instead, large larvae grow a robust pterygoid sutured with a fully ossified trapezoidal vomer and a short maxilla. Small larvae are readily distinguished by tooth count, morphology and arrangement from more advanced larvae. Age‐related features, irrespective of metamorphosis, include pedicellate teeth, morphological differentiation of parasphenoid, enlargement of the orbitosphenoid, distal expansion of columella, and loss of coronoid teeth. 相似文献
9.
Isabella Soreca Andrea Fagiolini Ellen Frank Bret H. Goodpaster David J. Kupfer 《Chronobiology international》2013,30(4):780-788
We explored whether obesity in patients with bipolar disorder is associated with their chronotype. A group of 29 patients with bipolar I disorder, not currently experiencing an affective episode, were assessed for total body fat, mood symptoms, and self-reported circadian chronotype and sleep quality. Chronotype explained 19% of the variance in body fat, after age, sex, mood state, and sleep quality were accounted for. This association suggested that evening chronotype patients have a higher percentage of total body fat. Evening chronotype could be a proxy for as yet unknown specific causes of the high rate of obesity and obesity-related diseases in bipolar disorder. 相似文献
10.
Gary M. Kupfer 《The Yale journal of biology and medicine》2013,86(4):491-497
Fanconi anemia (FA) is a fascinating, rare genetic disorder marked by congenital
defects, bone marrow failure, and cancer susceptibility. Research in recent
years has led to the elucidation of FA as a DNA repair disorder and involved
multiple pathways as well as having wide applicability to common cancers,
including breast, ovarian, and head and neck. This review will describe the
clinical aspects of FA as well as the current state of its molecular
pathophysiology. In particular, work from the Kupfer laboratory will be
described that demonstrates how the FA pathway interacts with multiple DNA
repair pathways, including the mismatch repair system and signal transduction
pathway of the DNA damage response. 相似文献