Improved telomere detection using a telomere repeat probe (TTAGGG)<sub>n</sub>generated by PCR

IJdo, Jacob W.; Wells, Richard A.; Baldini, Antonio; Reeders, Stephen T.

doi:10.1093/nar/19.17.4780

Cited by 545 publications

(366 citation statements)

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“…The fluorescence in situ hybridization (FISH) was performed according to Pinkel et al (1986), using 18S rDNA (Hatanaka and Galetti Junior, 2004), 5S rDNA (Martins et al, 2000), [TTAGGG] n and [GATA] n probes, which were amplified without template DNA through PCR reaction, as described by Ijdo et al (1991). The 18S rDNA probe was labeled with biotin-16-dUTP by nick translation according to the manufacturer's instruction (Biotin Nick Translation mix -Roche).…”

Section: Methodsmentioning

confidence: 99%

Karyotypic variation of Glanidium ribeiroi Haseman, 1911 (Siluriformes, Auchenipteridae) along the Iguazu river basin

et al. 2015

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The Iguazu river is a tributary of the left margin of the Paraná river, isolated from this basin about 22 million years ago with the appearance of the Iguazu Falls. The Iguazu river is characterized by high endemism due to two factors: its rugged topography and the old isolation caused by formation of the Iguazu Falls. This study analyzed cytogenetically a population of Glanidium ribeiroi collected in a region at the final stretch of this basin, by Giemsa staining, C-banding, impregnation by silver nitrate, and FISH with probes of 5S rDNA, 18S rDNA, telomeric sequence [TTAGGG] n , and [GATA] n repeats. The diploid number was equal to 58 chromosomes. The heterochromatin was present in the terminal region of almost all chromosomes. The Ag-NORs were simple and presented interstitially on the short arm of the submetacentric pair 14, which was confirmed by FISH with 18S rDNA probe. The 5S rDNA-FISH marked only the submetacentric pair 16 on the long arm in interstitial position. The FISH with [TTAGGG] n probe presented all telomeres labeled as expected, with an absence of Interstitial Telomeric Sequence (ITS). The repetitive [GATA] n sequence was dispersed throughout the genome, with preferential location in the terminal region of all chromosomes. The data obtained are discussed herein with other species of Auchenipteridae, and other previously analyzed populations of G. ribeiroi from the Iguazu river, verifying differences among these populations, which should be mainly related to the rugged topography of this basin.Keywords: geographical barriers, ribosomal DNA, Glanidium ribeiroi, Iguazu river. Variação cariotípica de Glanidium ribeiroi

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Section: Methodsmentioning

confidence: 99%

Karyotypic variation of Glanidium ribeiroi Haseman, 1911 (Siluriformes, Auchenipteridae) along the Iguazu river basin

et al. 2015

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“…Probes for detection of telomeric sequences ttaggg (n) were produced using the method published by Ijdo et al (1991b). Details of the protocol are given in the Supplementary File S3.…”

Section: Cytogenetic Analysismentioning

confidence: 99%

Independent fusions and recent origins of sex chromosomes in the evolution and diversification of glass knife fishes (Eigenmannia)

et al. 2010

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The genus Eigenmannia comprises several species groups that display a surprising variety of diploid chromosome numbers and sex-determining systems. In this study, hypotheses regarding phylogenetic relationships and karyotype evolution were investigated using a combination of molecular and cytogenetic methods. Phylogenetic relationships were analyzed for 11 cytotypes based on sequences from five mitochondrial DNA regions. Parsimony-based character mapping of sex chromosomes confirms previous suggestions of multiple origins of sex chromosomes. Molecular cytogenetic analyses involved chromosome painting using probes derived from whole sex chromosomes from two taxa that were hybridized to metaphases of their respective sister cytotypes. These analyses showed that a multiple XY system evolved recently (o7 mya) by fusion. Furthermore, one of the chromosomes that fused to form the neo-Y chromosome is fused independently to another chromosome in the sister cytotype. This may constitute an efficient post-mating barrier and might imply a direct function of sex chromosomes in the speciation processes in Eigenmannia. The other chromosomal sexdetermination system investigated is shown to have differentiated by an accumulation of heterochromatin on the X chromosome. This has occurred in the past 0.6 my, and is the most recent chromosomal sex-determining system described to date. These results show that the evolution of sexdetermining systems can proceed very rapidly.

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“…After complete digestion with HinfI and/or AluI, the genomic DNA was separated by 0.8% agarose gel electrophoresis (80 V, 6 h), was transferred to nylon membrane (Qiagen, Hilden, Germany), and was hybridized to P 32 -labeled probes of high molecular size telomeric repeats according to the manufacturer's protocol. The telomeric probe was generated in a template-free PCR by staggered annealing of telomere-oligonucleotides TEL1/2 (TTAGGG) 5 and TEL2/2 (CCCTAA) 5 using P 32 -dCTP in the nucleotide mix (Ijdo et al, 1991), which yielded doublestranded (TTAGGG/AATCCC)n repetitive fragments of up to 10 kBp length. The high molecular size telomeric amplification products were purified using the PCR purification kit (Qiagen).…”

Section: Analyses Of Telomere Lengthmentioning

confidence: 99%

The YeastTEL1Gene Partially Substitutes for HumanATMin Suppressing Hyperrecombination, Radiation-Induced Apoptosis and Telomere Shortening in A-T Cells

Fritz¹,

Friedl²,

Zwacka

et al. 2000

MBoC

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Homozygous mutations in the human ATM gene lead to a pleiotropic clinical phenotype of ataxia-telangiectasia (A-T) patients and correlating cellular deficiencies in cells derived from A-T donors. Saccharomyces cerevisiae tel1 mutants lacking Tel1p, which is the closest sequence homologue to the ATM protein, share some of the cellular defects with A-T. Through genetic complementation of A-T cells with the yeast TEL1 gene, we provide evidence that Tel1p can partially compensate for ATM in suppressing hyperrecombination, radiation-induced apoptosis, and telomere shortening. Complementation appears to be independent of p53 activation. The data provided suggest that TEL1 is a functional homologue of human ATM in yeast, and they help to elucidate different cellular and biochemical pathways in human cells regulated by the ATM protein.

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Improved telomere detection using a telomere repeat probe (TTAGGG)_ngenerated by PCR

Cited by 545 publications

References 2 publications

Karyotypic variation of Glanidium ribeiroi Haseman, 1911 (Siluriformes, Auchenipteridae) along the Iguazu river basin

Karyotypic variation of Glanidium ribeiroi Haseman, 1911 (Siluriformes, Auchenipteridae) along the Iguazu river basin

Independent fusions and recent origins of sex chromosomes in the evolution and diversification of glass knife fishes (Eigenmannia)

The YeastTEL1Gene Partially Substitutes for HumanATMin Suppressing Hyperrecombination, Radiation-Induced Apoptosis and Telomere Shortening in A-T Cells

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Improved telomere detection using a telomere repeat probe (TTAGGG)ngenerated by PCR

Cited by 545 publications

References 2 publications

Karyotypic variation of Glanidium ribeiroi Haseman, 1911 (Siluriformes, Auchenipteridae) along the Iguazu river basin

Karyotypic variation of Glanidium ribeiroi Haseman, 1911 (Siluriformes, Auchenipteridae) along the Iguazu river basin

Independent fusions and recent origins of sex chromosomes in the evolution and diversification of glass knife fishes (Eigenmannia)

The YeastTEL1Gene Partially Substitutes for HumanATMin Suppressing Hyperrecombination, Radiation-Induced Apoptosis and Telomere Shortening in A-T Cells

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Improved telomere detection using a telomere repeat probe (TTAGGG)_ngenerated by PCR