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81.
82.
Pavlos Neophytou Rolandos Constantinides Akis Lazarou Alkis Pierides C. Constantinou Deltas 《Human genetics》1996,98(4):437-442
Mutations in the PKD1 gene on the short arm of chromosome 16 account for 85%–90% of polycystic kidney disease patients in
the Caucasian population. After the recent characterization of the gene, we started a search for mutations in its 3′-end unique
portion in Cypriot patients, by using the method of single-strand conformation polymorphism (SSCP). In one large family, we
identified a nucleotide substitution at position 12 258 of the cDNA; this substitutes cysteine-4086 by a premature termination
codon (C4086X). It has been inherited by every affected family member but not by unaffected members, nor by patients from
13 other Cypriot families. A new polymerase chain reaction (PCR) primer has been designed to engineer a novel DdeI recognition site upon PCR amplification, thereby allowing easy detection of the mutation by PCR-restriction digestion. The
premature STOP codon is expected to remove 217 residues from the putative C-terminal intracellular domain of the gene product,
polycystin and thus identifies this part as being critical to the production of the disease phenotype, possibly by interfering
with the transmission of signals from the extracellular matrix to the cytoplasm. We also describe the identification of the
first polymorphism within the encoding region of the gene. It is at alanine 4091, which is encoded by either GCA or GCG. With
a heterozygosity of 35%, it should be extremely useful in informative families, especially because the gene lies in an unstable
region and is prone to rearrangements. This polymorphism is readily detectable by PCR-restriction digestion with Bsp1286I.
Received: 19 February 1996 / Revised: 20 April 1996 相似文献
83.
Phylogenetic analysis of slippage-like sequence variation in the V4 rRNA expansion segment in tiger beetles (Cicindelidae) 总被引:1,自引:1,他引:0
Sequence variation in the middle part of the small-subunit rRNA was studied
for representatives of the major groups in the family Cicindelidae
(Coleoptera). All taxa exhibited a much expanded segment in variable region
V4 compared to D. melanogaster. This expanded segment was not found in
other groups of beetles, including three taxa in the closely related
Carabidae. Secondary structure predictions indicate that the expanded
segment folds into a single stem-loop structure in all taxa. Despite its
structural conservation, the fragment differs strongly in primary sequence,
even between closely related sister taxa. Several features of these
sequences are consistent with slippage replication as the mechanism that
has generated this sequence variation: the level of internal sequence
repetition as measured by the relative simplicity factor (RSF), its
variation in length between close relatives, and the strong nucleotide bias
compared to the remainder of the gene. With few exceptions, there was also
a correlation between sequence length and the level of sequence repetition,
frequently interpreted as the result of slippage. Phylogenies inferred from
the expansion segment were not consistent with existing hypotheses from
other molecular data for the group. This indicates that DNA sequences in
this region are not homologous throughout the entire Cicindelidae, but it
leaves open the possibility that this expansion segment can be used for
phylogeny reconstruction within subgroups. The implications of a
phylogenetic approach to the understanding of slippage-like evolution are
discussed.
相似文献
84.
85.
Nimmy Mohan Sudheesh AP Nimmy Francis Richard Anderson Rakesh S. Laishram 《Nucleic acids research》2015,43(14):7005-7020
Star-PAP is a nuclear non-canonical poly(A) polymerase (PAP) that shows specificity toward mRNA targets. Star-PAP activity is stimulated by lipid messenger phosphatidyl inositol 4,5 bisphoshate (PI4,5P2) and is regulated by the associated Type I phosphatidylinositol-4-phosphate 5-kinase that synthesizes PI4,5P2 as well as protein kinases. These associated kinases act as coactivators of Star-PAP that regulates its activity and specificity toward mRNAs, yet the mechanism of control of these interactions are not defined. We identified a phosphorylated residue (serine 6, S6) on Star-PAP in the zinc finger region, the domain required for PIPKIα interaction. We show that S6 is phosphorylated by CKIα within the nucleus which is required for Star-PAP nuclear retention and interaction with PIPKIα. Unlike the CKIα mediated phosphorylation at the catalytic domain, Star-PAP S6 phosphorylation is insensitive to oxidative stress suggesting a signal mediated regulation of CKIα activity. S6 phosphorylation together with coactivator PIPKIα controlled select subset of Star-PAP target messages by regulating Star-PAP-mRNA association. Our results establish a novel role for phosphorylation in determining Star-PAP target mRNA specificity and regulation of 3′-end processing. 相似文献
86.
The critical micelle concentrations (CMCs) of palmitoyl-CoA/stearoyl-CoA and palmitoyl-CoA/oleoyl-CoA mixtures in 0.050 M KPi, pH 7.4, a buffer used in enzymatic studies, were determined by fluorescence. Mixed micelle solution theory, analogous to the thermodynamic treatment of vapor pressure, was applied to calculate monomer and micelle compositions. The behavior of the palmitoyl-CoA/stearoyl-CoA mixture is ideal, while the palmitoyl-CoA/oleoyl-CoA mixture, although not exhibiting ideal behavior, can be fitted reasonably well by nonideal theory. In both mixtures, selective micellization takes place and, unlike the case of pure fatty acyl-CoAs, above the CMC of the mixtures the concentration of molecules free in solution is strongly dependent upon total concentration. The information derived from the present physical studies becomes important in enzymatic studies with membrane-bound acyltransferases, where selectivity toward various fatty acyl donors, presented as binary mixtures, is frequently observed. 相似文献
87.
Modeling, optimization, and computer control of the cephalosporin C fermentation process 总被引:1,自引:0,他引:1
In this study, a cephalosporin C producing strain, Cephalosporium acremonium (ATCC 36225), was chosen to determine the optimal conditions that maximize antibiotic production in a mixed substrate of glucose and sucrose. A model for cell growth and cephalosporin C production at different pH and temperature was developed and the associated parameters were evaluated experimentally. Pontryagin's maximum principle, in conjunction with the model, was used to predict the optimal temperature and pH control profiles to maximize the production of antibiotic. 相似文献
88.
Simultaneous saccharification and fermentation of straw to ethanol using the thermotolerant yeast strain Kluyveromyces marxianus imb3 总被引:1,自引:0,他引:1
The thermotolerant, ethanol-producing yeast strain Kluyveromyces marxianus IMB3 was grown at 45°C on media containing 2, 4 and 6 % (w/v) pulverized barley straw and supplemented with 2% (v/v) cellulase. Maximum ethanol concentrations produced were 2, 3 and 3.6g/l, respectively. When the pulverized straw was replaced by NaOH pretreated straw (at 2, 4 and 6% (w/v); based on original untreated straw), ethanol concentrations increased to maxima of 3.9, 8, and 12g/l, respectively. The ethanol yields amount to 20g ethanol from 100g of straw. 相似文献
89.
Geraldine Spies-karotkin Spiros M. Constantinides 《Molecular and cellular biochemistry》1978,21(3):153-160
Summary Partially purified flounder muscle (Pseudopleuronectus americanus) glyceraldehyde 3-phosphate dehydrogenase was immobilized on cyanogen bromide-activated Sepharose. The catalytic properties of the immobilized preparation were studied to determine if immobilization alters the kinetic properties of the native holoenzyme. The results indicate that the pH activity profile of immobilized glyceraldehyde 3-phosphate dehydrogenase did not differ from that of the native enzyme. The Michaelis constants (Km) for NAD and glyceraldehyde 3-phosphate were somewhat altered. The enzyme stability toward various inactivation treatments in the presence and absence of NAD was characterized and compared to that of he native enzyme. When either form of the enzyme was incubated with urea at concentrations greater than 2m, inactivation occurred very rapidly. Incubation in 0.1% trypsin for 60 minutes decreased the activity of immobilized glyceraldehyde 3-phosphate dehydrogenase by 45% and of the native soluble enzyme by 70%. The immobilized enzyme also exhibited considerably more stability than the native soluble enzyme when exposed to a temperature of 50° or to 20 mm ATP. In all cases NAD either greatly reduced the rate of inactivation or completely protected the enzyme from inactivation. 相似文献
90.
Evolutionary relatedness of some primate models of Plasmodium 总被引:1,自引:0,他引:1
Primate--and, specifically, monkey--malaria infections are commonly used
for understanding the pathology of and immune response to the human disease
because they are thought to resemble most closely the host-parasite
relationship found in humans. Plasmodium cynomolgi is used extensively as a
model for the human parasite, P. vivax, and P. knowlesi is used primarily
as a model for the development of erythrocytic-stage vaccines. Both of
these simian parasites can naturally infect man, resulting in mildly
symptomatic episodes of the disease. The phylogenetic relationship between
these two simian parasites and previously characterized Plasmodium species,
including P. vivax, was examined by comparison of the asexually expressed
small- subunit ribosomal RNA genes. Our analysis confirmed that P. vivax is
most closely related to P. cynomolgi and that it remains an appropriate
model of the human pathogen. Furthermore, with P. knowlesi and P. fragile,
these two species form a group of closely related species, distant from
other Plasmodium species. What is considered to be the most ancient of the
human malaria pathogens, P. malariae, was also included in the analysis and
does not group at all with other simian or human parasites.
相似文献