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排序方式: 共有790条查询结果,搜索用时 15 毫秒
91.
Lamarche F Mével M Montier T Burel-Deschamps L Giamarchi P Tripier R Delépine P Le Gall T Cartier D Lehn P Jaffrès PA Clément JC 《Bioconjugate chemistry》2007,18(5):1575-1582
The DNA compacting properties of polyamines (especially spermine) are well-known, hence the use of spermine as the cationic part in several synthetic DNA carriers. Here, we describe the synthesis of modified spermines, with a "lipophosphoramidate" as the lipidic part, and their use for efficient in vitro transfection. Physicochemical measurements (particle size, zeta potentials, pKa determination) and gel retardation assays were also performed. Theoretical membrane-disrupting ability was established by FRET. Taken together, our results indicate that lipophosphoramidates constitute an interesting alternative to "classical" lipidic parts of cationic lipids used as DNA carriers. 相似文献
92.
Rescue and production of vaccine and therapeutic adenovirus vectors expressing inhibitory transgenes
Gall JG Lizonova A EttyReddy D McVey D Zuber M Kovesdi I Aughtman B King CR Brough DE 《Molecular biotechnology》2007,35(3):263-273
Expression of certain transgenes from an adenovirus vector can be deleterious to its own replication. This can result in the
inhibition of virus rescue, reduced viral yields, or, in the worst case, make it impossible to construct a vector expressing
the inhibiting transgene product. A gene regulation system based on the tet operon was used to allow the rescue and efficient growth of adenovectors that express transgenes to high levels. A key advantage
to this system is that repression of transgene expression is mediated by the packaging cell line, thus, expression of regulatory
products from the adenovector are not required. This provides a simple, broadly applicable system wherein transgene repression
is constitutive during vector rescue and growth and there is no effect on adenovector-mediated expression of gene products
in transduced cells. Several high-level expression vectors based on first- and second-generation adenovectors were rescued
and produced to high titer that otherwise could not be grown. Yields of adenovectors expressing inhibitory transgene products
were increased, and the overgrowth of cultures by adenovectors with nonfunctional expression cassettes was prevented. The
gene regulation system is a significant advancement for the development of adenovirus vectors for vaccine and other gene transfer
applications. 相似文献
93.
94.
Megan D. Gall Walter Wilczynski 《Proceedings. Biological sciences / The Royal Society》2015,282(1808)
We investigated whether hearing advertisement calls over several nights, as happens in natural frog choruses, modified the responses of the peripheral auditory system in the green treefrog, Hyla cinerea. Using auditory evoked potentials (AEP), we found that exposure to 10 nights of a simulated male chorus lowered auditory thresholds in males and females, while exposure to random tones had no effect in males, but did result in lower thresholds in females. The threshold change was larger at the lower frequencies stimulating the amphibian papilla than at higher frequencies stimulating the basilar papilla. Suprathreshold responses to tonal stimuli were assessed for two peaks in the AEP recordings. For the peak P1 (assessed for 0.8–1.25 kHz), peak amplitude increased following chorus exposure. For peak P2 (assessed for 2–4 kHz), peak amplitude decreased at frequencies between 2.5 and 4.0 kHz, but remained unaltered at 2.0 kHz. Our results show for the first time, to our knowledge, that hearing dynamic social stimuli, like frog choruses, can alter the responses of the auditory periphery in a way that could enhance the detection of and response to conspecific acoustic communication signals. 相似文献
95.
Alexey?V. Krasnoslobodtsev Yuliang Zhang Ekaterina Viazovkina Alexander Gall Chad Bertagni Yuri?L. Lyubchenko 《Biophysical journal》2015,108(9):2333-2339
Immobilization is a key step involved in probing molecular interactions using single-molecule force spectroscopy methods, including atomic force microscopy (AFM). To our knowledge, we describe a novel approach termed flexible nanoarray (FNA) in which the interaction between the two internally immobilized amyloid β peptides is measured by pulling of the tether. The FNA tether was synthesized with nonnucleotide phosphoramidite monomers using the DNA synthesis chemistry. The two anchoring points for immobilization of the peptides inside the tether were incorporated at defined distances between them and from the ends of the polymer. Decamers of amyloid β peptide capable of dimer formation were selected as a test system. The formation of the peptide dimers was verified by AFM force spectroscopy by pulling the tether at the ends. In these experiments, the thiolated end of the FNA tether was covalently immobilized on the AFM substrate functionalized with maleimide. The other end of the FNA tether was functionalized with biotin to form a noncovalent link with the streptavidin functionalized AFM tip during the approach stage. The dimers’ rupture fingerprint was unambiguously identified on the force curves by its position and the force value. The FNA design allowed reversible experiments in which the monomers were allowed to associate after the rupture of the dimers by performing the approach stage before the rupture of the biotin-streptavidin link. This suggests that the FNA technique is capable of analyzing multiple intermolecular interactions in the same molecular complex. The computational analysis showed that the tethered peptides assemble into the same dimer structure as that formed by nontethered peptides, suggesting that the FNA tether has the necessary flexibility to enable assembly of the dimer even during the course of the force spectroscopy experiment. 相似文献
96.
Thorsten Maretzky Astrid Evers Sylvain Le Gall Rolake O. Alabi Nancy Speck Karina Reiss Carl P. Blobel 《The Journal of biological chemistry》2015,290(12):7416-7425
The membrane-anchored metalloproteinase a disintegrin and metalloprotease 10 (ADAM10) is required for shedding of membrane proteins such as EGF, betacellulin, the amyloid precursor protein, and CD23 from cells. ADAM10 is constitutively active and can be rapidly and post-translationally enhanced by several stimuli, yet little is known about the underlying mechanism. Here, we use ADAM10-deficient cells transfected with wild type or mutant ADAM10 to address the role of its cytoplasmic and transmembrane domain in regulating ADAM10-dependent protein ectodomain shedding. We report that the cytoplasmic domain of ADAM10 negatively regulates its constitutive activity through an ER retention motif but is dispensable for its stimulated activity. However, chimeras with the extracellular domain of ADAM10 and the transmembrane domain of ADAM17 with or without the cytoplasmic domain of ADAM17 show reduced stimulated shedding of the ADAM10 substrate betacellulin, whereas the ionomycin-stimulated shedding of the ADAM17 substrates CD62-L and TGFα is not affected. Moreover, we show that influx of extracellular calcium activates ADAM10 but is not essential for its activation by APMA and BzATP. Finally, the rapid stimulation of ADAM10 is not significantly affected by incubation with proprotein convertase inhibitors for up to 8 h, arguing against a major role of increased prodomain removal in the rapid stimulation of ADAM10. Thus, the cytoplasmic domain of ADAM10 negatively influences constitutive shedding through an ER retention motif, whereas the cytoplasmic domain and prodomain processing are not required for the rapid activation of ADAM10-dependent shedding events. 相似文献
97.
Gallè F Sanguinetti M Colella G Di Onofrio V Torelli R Rossano F Liguori G 《The new microbiologica》2011,34(4):379-389
Recurrent oral candidosis is a common problem in immunocompromised patients, and it is frequently triggered by resistance induced by antifungal treatment. Knowledge of the mechanisms by which the yeast persists in the host could allow the management of this type of infection. This study used electrophoretic karyotyping and restriction fragment length polymorphism based on the use of 27A probe to study 12 pairs of Candida albicans isolates from patients with recurrent candidosis to distinguish new infections from relapses caused by the same strain responsible for the first episode. Subsequently, RT-PCR was used to evaluate expression of CDR1, CDR2 and MDR1 genes, which are involved in C. albicans azole resistance, in the three pairs that consisted of variants of the same strain. Restriction polymorphism resulted in better discrimination than with karyotyping in defining differences between strains. In one case, RT-PCR allowed us to identify deregulation of efflux pump genes as the possible underlying mechanism in recurrent candidosis. The techniques employed resulted effective for the characterization of recurrent oral candidosis. Broader analysis could help to control better these infections and choose adequate therapy. 相似文献
98.
Bittner L Payri CE Maneveldt GW Couloux A Cruaud C de Reviers B Le Gall L 《Molecular phylogenetics and evolution》2011,61(3):697-713
Systematics of the red algal order Corallinales has a long and convoluted history. In the present study, molecular approaches were used to assess the phylogenetic relationships based on the analyses of two datasets: a large dataset of SSU sequences including mainly sequences from GenBank; and a combined dataset including four molecular markers (two nuclear: SSU, LSU; one plastidial: psbA; and one mitochondrial: COI). Phylogenetic analyses of both datasets re-affirmed the monophyly of the Corallinales as well as the two families (Corallinaceae and Hapalidiaceae) currently recognized within the order. Three of the four subfamilies of the Corallinaceae (Corallinoideae, Lithophylloideae, Metagoniolithoideae) were also resolved as a monophyletic lineage whereas members of the Mastophoroideae were resolved as four distinct lineages. We therefore propose to restrict the Mastophoroideae to the genera Mastophora, Metamastophora, and possibly Lithoporella in the aim of rendering this subfamily monophyletic. In addition, our phylogenies resolved the genus Hydrolithon in two unrelated lineages, one containing the generitype Hydrolithon reinboldii and the second containing Hydrolithon onkodes, which used to be the generitype of the now defunct genus Porolithon. We therefore propose to resurrect the genus Porolithon for the second lineage encompassing those species with primarily monomerous thalli, and trichocyte arrangements in large pustulate horizontal rows. Moreover, our phylogenetic analyses revealed the presence of cryptic diversity in several taxa, shedding light on the need for further studies to better circumscribe species frontiers within the diverse order Corallinales, especially in the genera Mesophyllum and Neogoniolithon. 相似文献
99.
Thirant C Varlet P Lipecka J Le Gall M Broussard C Chafey P Studler JM Lacombe J Lions S Guillaudeau A Camoin L Daumas-Duport C Junier MP Chneiweiss H 《Proteomics》2011,11(21):4139-4154
Gliomas are primary tumors of the human central nervous system with unknown mechanisms of progression. Isocitrate dehydrogenase-1 (IDH1) mutation is frequent in diffuse gliomas such as oligodendrogliomas. To gain insights into the physiopathology of oligodendrogliomas that have a better prognosis than other diffuse gliomas, we combined microdissection, 2-D DIGE and MS/MS focusing on proteome alterations associated with IDH1 mutation. We first compared tumor tissues (TT) and minimally infiltrated parenchymal tissues (MIT) of four IDH1-mutated oligodendrogliomas to verify whether proteins specific to oligodendroglioma tumor cells could be identified from one patient to another. This study resulted in identification of 68 differentially expressed proteins, with functions related to growth of tumor cells in a nervous parenchyma. We then looked for proteins distinctly expressed in TT harboring either mutant (oligodendrogliomas, n=4) or wild-type IDH1 (oligodendroglial component of malignant glio-neuronal tumors, n=4). This second analysis resulted in identification of distinct proteome patterns composed of 42 proteins. Oligodendrogliomas with a mutant IDH1 had noteworthy enhanced expression of enzymes controlling aerobic glycolysis and detoxification, and anti-apoptosis proteins. In addition, the mutant IDH1 migrated differently from the wild-type IDH1 form. Comparative proteomic analysis might thus be suitable to identify proteome alterations associated with a well-defined mutation. 相似文献
100.
Plant virus RNAs. Coordinated recruitment of conserved host functions by (+) ssRNA viruses during early infection events
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Thivierge K Nicaise V Dufresne PJ Cotton S Laliberté JF Le Gall O Fortin MG 《Plant physiology》2005,138(4):1822-1827
Positive-sense single-stranded RNA viruses have developed strategies to exploit cellular resources at the expense of host mRNAs. The genomes of these viruses display a variety of structures at their 5' and 3' ends that differentiate them from cellular mRNAs. Despite this structural diversity, viral RNAs are still circularized by juxtaposition of their 5' and 3' ends, similar to the process used by cellular mRNAs. Also reminiscent of the mechanisms used by host mRNAs, translation of viral RNAs involves the recruitment of translation initiation factors. However, the roles played by these factors likely differ from those played by cellular mRNAs. In keeping with the general parsimony typical of RNA viruses, these host factors also participate in viral RNA replication. However, the dual use of host factors requires that viral RNA template utilization be regulated to avoid conflict between replication and translation. The molecular composition of the large ribonucleoprotein complexes that form the viral RNA replication and translation machineries likely evolves over the course of infection to allow for switching template use from translation to replication. 相似文献