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Physical mapping of genes by fluorescence in situ hybridization (FISH) seems to be out of fashion in species whose assembled genome sequences are available. However, in this work we evidence the existence of errors in gene location in the Btau_4.0 assembly. We show that DFNA5 and CHCHD6 genes are located on BTA4 and BTA22, respectively, instead of BTA10 and BTA3, as displayed by Btau_4.0. This report emphasizes the need to verify the data on physical localization of genes in the cattle genome (at least by taking into account comparative data reported in available papers) and the need to improve the cattle genome assembly. Our results indicate that FISH mapping in cattle is still useful.
We report on a phenotypically normal man with infertility, whose 47,XY,+mar karyotype was studied by spectral karyotyping (SKY) and fluorescence in situ hybridization (FISH) using a chromosome-15-specific probe (LSI SNRPN). By these techniques, the marker chromosome was identified as a small inv dup (15). Possible causes for male infertility in this case are discussed.
Prokaryotes from different phylogenetic groups were studied in surface microlayer (SM, up to 100 μm) and subsurface water (SW – 20 cm) in a eutrophic lake over three months (July, August, and October). The abundance of prokaryotes was determined by epifluorescence microscopy after DAPI staining, and phylogenetic diversity was determined by fluorescence in situ hybridization (FISH) with group-specific, fluorescently labeled oligonucleotide probes. In SW bacteria made up most of the entire community of DAPI-stained microorganisms (54-69%) and in SM bacteria made up only 33-44% of DAPI-stained microorganisms. Archaea corresponded to a small fraction of both bacterioneuston and bacterioplankton. The counts of Archaea and bacteria were significantly higher in SW than in SM. Among all proteobacteria included in the research, γ-proteobacteria represented the most abundant fraction: 42-72% in SM and 39-61% in SW. Statistical analysis revealed that the abundance of γ-proteobacteria is positively correlated with temperature and with dissolved oxygen. β-proteobacteria were the least abundant fraction.
Fluorescent in situ hybridization (FISH) was performed in 76 patients referred to our department because of intellectual disability and dysmorphic features that can be related to subtelomeric microaberrations. In all the patients, conventional cytogenetic methods revealed normal karyotype. Four (5.3%) subtelomeric rearrangements were detected by FISH: 2 subtelomeric 1p36 deletions, an unbalanced translocation involving chromosomes 1 and 12 with 1p36 deletion, and a de novo balanced translocation involving chromosomes 19 and 22. Thus, 3 cases of 1p36 subtelomeric deletion were found (3.95%). To confirm subtelomeric rearrangements in 2 patients, comparative genomic hybridization (CGH) was applied. Moreover, 3 cases of polymorphism without phenotypic effects were found: in 2 patients, the polymorphism involved the long arm of chromosome 2 (maternal derivative in both patients), while in the third patient, a polymorphism of the long arm of chromosome 7 was diagnosed. The latter polymorphism was also found in the patient's mother and grandfather.
We report on a 13-month-old girl showing dysmorphic features and a delay in psychomotor development. She was diagnosed with a balanced de novo translocation 46,X,t(X; 13)(p 11,2; p 13) and non-random inactivation of the X chromosome. FISH analysis, employing the X chromosome centromere and XIST-region-specific probes, showed that the XIST locus was not involved in the translocation. Selective inactivation of paternal X, which was involved in translocation, was revealed by the HUMARA assay. The pattern of methylation of 5 genes located within Xp, which are normally silenced on an inactive X chromosome, corresponded to an active (unmethylated) X chromosome. These results revealed that in our proband the X chromosome involved in translocation (Xt) was preferentially inactivated. However, genes located on the translocated Xp did not include XIST. This resulted in functional Xp disomy, which most probably accounts for the abnormal phenotype in our patient.
Fluorescence and genomic in situ hybridization (FISH and GISH) methods were used for discrimination of Brassica genomes. The three diploid and three allotetraploid species of Brassica, known as the "U-triangle," represent an attractive model for molecular and cytologieal analysis of genome changes during phylogeny in the genus Brassica. The use of genomic DNA probes enabled unambiguous discrimination of the ancestral genomes in B. juncea and B. carinata, and was only partially successful in B. napus. GISH signals in all genomes were localized predominantly in pericentromeric regions of chromosomes. Simultaneous application of genomic and ribosomal DNA probes in multicolor GISH and FISH allowed identification of a significant number of chromosomes in the B. juncea complement. The study also revealed that species of Brassica possess Arabidopsis-type telomeric repeats which in all genomes occupied exclusively terminal, that is, telomeric, locations of chromosomes.
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