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41.
Mitochondrial DNA and bindin gene sequence evolution among allopatric species of the sea urchin genus Arbacia 总被引:3,自引:1,他引:2
Sea urchins of the genus Arbacia (order Stirodonta) have discontinuous
allopatric distributions ranging over thousands of kilometers.
Mitochondrial DNA (mtDNA) sequences were used to reconstruct phylogenetic
relationships of four Arbacia species and their geographic populations.
There is little evidence of genetic structuring of populations within
species, except in two cases at range extremes. The mtDNA sequence
differentiation between species suggests that divergence occurred about 4-9
MYA. Gene sequences encoding the sperm protein bindin and its intron were
obtained and compared with the mtDNA phylogeny. Sea urchins among the
well-studied echinoid order Camarodonta, with degrees of mtDNA divergence
similar to those of Arbacia species, are known to have remarkable variation
in bindin. However, in Arbacia, little variation in deduced amino acid
sequences of bindin was found, indicating that purifying selection acts on
the protein. In contrast, bindin intron sequences showed much
differentiation, including numerous insertion/deletions. Fertilization
experiments performed between a divergent pair of Arbacia species from the
Atlantic and Pacific Oceans revealed no evidence of blocks to gamete
recognition. In Arbacia, fertilization specificities may have evolved
relatively slowly as a result of extensive gene flow within species,
greater functional constraint on the bindin polypeptide, or reduced
selective pressure for species recognition in singly occurring species.
相似文献
42.
Catimel B; Scott AM; Lee FT; Hanai N; Ritter G; Welt S; Old LJ; Burgess AW; Nice EC 《Glycobiology》1998,8(9):927-938
We describe a novel immobilization technique to investigate interactions
between immobilized gangliosides (GD3, GM1, and GM2) and their respective
antibodies, antibody fragments, or binding partners using an optical
biosensor. Immobilization was performed by direct injection onto a
carboxymethyldextran sensor chip and did not require derivatization of the
sensor surface or the ganglioside. The ganglioside appeared to bind to the
sensor surface by hydrophobic interaction, leaving the carbohydrate epitope
available for antibody or, in the case of GM1, cholera toxin binding. The
carboxyl group of the dextran chains on the sensor surface did not appear
to be involved in the immobilization as evidenced by equivalent levels of
immobilization following conversion of the carboxyl groups into acyl amino
esters, but rather the dextran layer provided a hydrophilic coverage of the
sensor chip which was essential to prevent nonspecific binding. This
technique gave better reactivity and specificity for anti- ganglioside
monoclonal antibodies (anti-GD3: KM871, KM641, R24; and anti-GM2: KM966)
than immobilization by hydrophobic interaction onto a gold sensor surface
or photoactivated cross-linking onto carboxymethydextran. This rapid
immobilization procedure has facilitated detailed kinetic analysis of
ganglioside/antibody interactions, with the surface remaining viable for a
large number of cycles (>125). Kinetic constants were determined from
the biosensor data using linear regression, nonlinear least squares and
equilibrium analysis. The values of kd, ka, and KAobtained by nonlinear
analysis (KAKM871 = 1.05, KM641 = 1.66, R24 = 0.14, and KM966 = 0.65 x
10(7) M- 1) were essentially independent of concentration and showed good
agreement with data obtained by other analytical methods.
相似文献
43.
HIV-particles in spermatozoa of patients with AIDS and their transfer into the oocyte 总被引:11,自引:1,他引:10
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B Baccetti A Benedetto AG Burrini G Collodel EC Ceccarini N Crisa A Di Caro M Estenoz AR Garbuglia A Massacesi P Piomboni T Renieri D Solazzo 《The Journal of cell biology》1994,127(4):903-914
By immunocytochemistry and in situ hybridization at the electron microscopy level, and by the PCR technique, we have shown that HIV-1 binds and enters normal sperm; that viral particles, their antigens, and nucleic acid are present in sperm from HIV-1 infected men; and that such sperm can transfer HIV-1 like particles to normal human oocytes. We also present evidence that a galactosylceramide-like compound is present on the sperm membrane and could function as an alternative receptor for HIV. 相似文献
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47.
In 1997–99 the occurrence ofFusarium spp. on winter wheat and the contamination with mycotoxins was investigated at three locations in the Rhineland, Germany. All cultivation methods investigated had an effect on the level ofFusarium infection, however, rainfall during flowering was the most important factor. The choice of cultivar and soil cultivation proved to be the most promising tools to reduce head scab severity and mycotoxin contamination. 相似文献
48.
Essers J Theil AF Baldeyron C van Cappellen WA Houtsmuller AB Kanaar R Vermeulen W 《Molecular and cellular biology》2005,25(21):9350-9359
The DNA polymerase processivity factor proliferating cell nuclear antigen (PCNA) is central to both DNA replication and repair. The ring-shaped homotrimeric PCNA encircles and slides along double-stranded DNA, acting as a "sliding clamp" that localizes proteins to DNA. We determined the behavior of green fluorescent protein-tagged human PCNA (GFP-hPCNA) in living cells to analyze its different engagements in DNA replication and repair. Photobleaching and tracking of replication foci revealed a dynamic equilibrium between two kinetic pools of PCNA, i.e., bound to replication foci and as a free mobile fraction. To simultaneously monitor PCNA action in DNA replication and repair, we locally inflicted UV-induced DNA damage. A surprisingly longer residence time of PCNA at damaged areas than at replication foci was observed. Using DNA repair mutants, we showed that the initial recruitment of PCNA to damaged sites was dependent on nucleotide excision repair. Local accumulation of PCNA at damaged regions was observed during all cell cycle stages but temporarily disappeared during early S phase. The reappearance of PCNA accumulation in discrete foci at later stages of S phase likely reflects engagements of PCNA in distinct genome maintenance processes dealing with stalled replication forks, such as translesion synthesis (TLS). Using a ubiquitination mutant of GFP-hPCNA that is unable to participate in TLS, we noticed a significantly shorter residence time in damaged areas. Our results show that changes in the position of PCNA result from de novo assembly of freely mobile replication factors in the nucleoplasmic pool and indicate different binding affinities for PCNA in DNA replication and repair. 相似文献
49.
50.
Hybrids monosomal for human chromosome 5 reveal the presence of a spinal muscular atrophy (SMA) carrier with two SMN1 copies on one chromosome 总被引:3,自引:0,他引:3
Mailman MD Hemingway T Darsey RL Glasure CE Huang Y Chadwick RB Heinz JW Papp AC Snyder PJ Sedra MS Schafer RW Abuelo DN Reich EW Theil KS Burghes AH de la Chapelle A Prior TW 《Human genetics》2001,108(2):109-115
We have analyzed the survival motor neuron gene (SMN1) dosage in 100 parents of children with homozygous SMN1 deletions. Of these parents, 96 (96%) demonstrated the expected one-copy SMN1 carrier genotype. However, four parents (4%) were observed to have a normal two-copy SMN1 dosage. The presence of two intact SMN1 genes in the parent of an affected child indicates either the occurrence of a de novo mutation event or a situation in which one chromosome has two copies of SMN1, whereas the other is null. We have separated individual chromosomes from two of these parents with two-copy SMN1 dosage by somatic cell hybridization and have employed a modified quantitative dosage assay to provide direct evidence that one parent is a two-copy/ zero-copy SMN1 carrier, whereas the other parent had an affected child as the result of a de novo mutation. These findings are important for assessing the recurrence risk of parents of children with spinal muscular atrophy and for providing accurate family counseling. 相似文献