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81.
Padlock oligonucleotides as a tool for labeling superhelical DNA   总被引:2,自引:1,他引:1       下载免费PDF全文
Labeling of a covalently closed circular double-stranded DNA was achieved using a so-called ‘padlock oligonucleotide’. The oligonucleotide was targeted to a sequence which is present in the replication origin of phage f1 and thus in numerous commonly used plasmids. After winding around the double-stranded target DNA sequence by ligand-induced triple helix formation, a biotinylated oligonucleotide was circularized using T4 DNA ligase and in this way became catenated to the plasmid. A gel shift assay was developed to measure the extent of plasmid modification by the padlock oligonucleotide. A similar assay showed that a modified supercoiled plasmid was capable of binding one streptavidin molecule thanks to the biotinylated oligonucleotide and that this binding was quantitative. The catenated complex was visualized by electron and atomic force microscopies using streptavidin conjugates or single strand-binding proteins as protein tags for the padlock oligonucleotide. This method provides a versatile tool for plasmid functionalization which offers new perspectives in the physical study of supercoiled DNA and in the development of improved vectors for gene therapy.  相似文献   
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Analysis of surface-spread synaptonemal complexes of zygotene and pachytene spermatocytes was carried out on a human male carrier of a pericentric inversion of chromosome 21 ascertained after four miscarriages. The synaptic behavior of the bivalent, which could be unambiguously identified by its nonaligned kinetochores, was analyzed. All zygotene and pachytene spermatocytes had 22 linearly paired autosomal bivalents, with apparently normal synaptonemal complexes, and no evidence of a loop configuration in the 50 cells analyzed. According to the XY type (classification of Solari), the cells were distributed across zygotene and pachytene stages, not exclusively in the late pachytene to which adjustment is conventionally thought to be confined. It is suggested that inverted segments heterosynapse at early pachytene, without previous homosynapsis. It is expected that this meiotic process leads to failure of crossing-over, reduces the production of unbalanced gametes, and the risk of recombinant offspring, but can increase the incidence of aneuploidy as a result of nondisjunction during meiosis I (a frequent cause of pregnancy wastage).  相似文献   
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Xavier Arnan  Alan N. Andersen  Heloise Gibb  Catherine L. Parr  Nathan J. Sanders  Robert R. Dunn  Elena Angulo  Fabricio B. Baccaro  Tom R. Bishop  Raphaël Boulay  Cristina Castracani  Xim Cerdá  Israel Del Toro  Thibaut Delsinne  David A. Donoso  Emilie K. Elten  Tom M. Fayle  Matthew C. Fitzpatrick  Crisanto Gómez  Donato A. Grasso  Blair F. Grossman  Benoit Guénard  Nihara Gunawardene  Brian Heterick  Benjamin D. Hoffmann  Milan Janda  Clinton N. Jenkins  Petr Klimes  Lori Lach  Thomas Laeger  Maurice Leponce  Andrea Lucky  Jonathan Majer  Sean Menke  Dirk Mezger  Alessandra Mori  Jimmy Moses  Thinandavha Caswell Munyai  Omid Paknia  Martin Pfeiffer  Stacy M. Philpott  Jorge L.P. Souza  Melanie Tista  Heraldo L. Vasconcelos  Javier Retana 《Global Change Biology》2018,24(10):4614-4625
The relationship between levels of dominance and species richness is highly contentious, especially in ant communities. The dominance‐impoverishment rule states that high levels of dominance only occur in species‐poor communities, but there appear to be many cases of high levels of dominance in highly diverse communities. The extent to which dominant species limit local richness through competitive exclusion remains unclear, but such exclusion appears more apparent for non‐native rather than native dominant species. Here we perform the first global analysis of the relationship between behavioral dominance and species richness. We used data from 1,293 local assemblages of ground‐dwelling ants distributed across five continents to document the generality of the dominance‐impoverishment rule, and to identify the biotic and abiotic conditions under which it does and does not apply. We found that the behavioral dominance–diversity relationship varies greatly, and depends on whether dominant species are native or non‐native, whether dominance is considered as occurrence or relative abundance, and on variation in mean annual temperature. There were declines in diversity with increasing dominance in invaded communities, but diversity increased with increasing dominance in native communities. These patterns occur along the global temperature gradient. However, positive and negative relationships are strongest in the hottest sites. We also found that climate regulates the degree of behavioral dominance, but differently from how it shapes species richness. Our findings imply that, despite strong competitive interactions among ants, competitive exclusion is not a major driver of local richness in native ant communities. Although the dominance‐impoverishment rule applies to invaded communities, we propose an alternative dominance‐diversification rule for native communities.  相似文献   
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Ansel M. and Thibaut M. 1973. Value of the specific distinction between Ascaris lumbricoïdes Linné 1758 and Ascaris suum Goeze 1782. International Journal for Parasitology3: 317–319. Study by scanning electron microscopy of heads of Ascaris lumbricoides Linné 1758 and Ascaris suum Goeze 1782 showed typical distinctive characteristics between these two species. There are differences in the appearance of the rows of denticules and the shapes of the lips.  相似文献   
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Mutations in the Ras oncogene are one of the most frequent events in human cancer. Although Ras regulates numerous growth-promoting pathways to drive transformation, it can paradoxically promote an irreversible cell cycle arrest known as oncogene-induced senescence. Although senescence has clearly been implicated as a major defense mechanism against tumorigenesis, the mechanisms by which Ras can promote such a senescent phenotype remain poorly defined. We have shown recently that the Ras death effector NORE1A plays a critical role in promoting Ras-induced senescence and connects Ras to the regulation of the p53 tumor suppressor. We now show that NORE1A also connects Ras to the regulation of a second major prosenescent tumor suppressor, the retinoblastoma (Rb) protein. We show that Ras induces the formation of a complex between NORE1A and the phosphatase PP1A, promoting the activation of the Rb tumor suppressor by dephosphorylation. Furthermore, suppression of Rb reduces NORE1A senescence activity. These results, together with our previous findings, suggest that NORE1A acts as a critical tumor suppressor node, linking Ras to both the p53 and the Rb pathways to drive senescence.  相似文献   
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