Restriction endonucleases in the study of eukaryotic chromosomes

López‐Fernández, Carmen; Gosálvez, Jaime; Ferrucci, Luigi; Mezzannotte, R.

doi:10.1007/bf00126232

Cited by 39 publications

(10 citation statements)

References 92 publications

(5 reference statements)

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“…In fact, the paracentromeric bands in S. sepsoides are Alu-l-resistant, whereas in C. chalcides they are digested by this endonuclease. On the other hand, the heterochromatin of C. ocellatus, like that of S. sepsoides, is Alu-1-resistant, thus suggesting either a similar composition of the satellite-like DNAs contained in the heterochromatin or a different composition coupled with a similar condensation pattern (Lopez-Fernandez et al, 1991). A detailed analysis of the satellite-like DNAs should provide resolution of this ques- tion. The karyological data for the species usually ascribed to the two genera Chalcides and Sphenops strongly suggest that they belong to the same natural taxon.…”

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confidence: 97%

Karyological Comparison of Sphenops sepsoides, Chalcides chalcides, and C. ocellatus (Reptilia: Scincidae): Taxonomic Implications

Caputo¹,

Odierna²,

Aprea³

1993

Copeia

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confidence: 97%

Karyological Comparison of Sphenops sepsoides, Chalcides chalcides, and C. ocellatus (Reptilia: Scincidae): Taxonomic Implications

Caputo¹,

Odierna²,

Aprea³

1993

Copeia

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“…In the present paper we present a modified in situ random primer extension procedure on exonuclease III treated chromosomes to show that there are exceptions to items (2) and (3) mentioned above. In fact, in the model we have tested, R bands coexist with labelled constitutive heterochromatin in chromosomes 1, 9, 16, and Yq.…”

mentioning

confidence: 94%

“…In the present paper we present a modified in situ random primer extension procedure on exonuclease III treated chromosomes to show that there are exceptions to items (2) and (3) Control slides were denatured in 70% formamide/2 x SSC at 70°C for three minutes, dehydrated in an increasing ethanol series, and incubated with the same probe. All slides were washed in 50% formamide/2 x SCC, pH7, at 43°C for 15 minutes and in 2 x SSC, pH7, at 37°C for eight minutes.…”

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confidence: 99%

Difference in constitutive heterochromatin behaviour between human amniocytes and lymphocytes detected by a sequential in situ exonuclease III digestion-random primer extension procedure.

Fernández

Campos

López‐Fernández

et al. 1995

Journal of Medical Genetics

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Fixed chromosomes from human amniotic fluid cells and peripheral blood lymphocytes were digested in situ with exonuclease III and the single stranded DNA obtained was used as template for an in situ random primer extension. Under these conditions an R banding pattern, more evident in lymphocytes than in amniocytes, was obtained. Nevertheless, constitutive heterochromatin of chromosomes 1, 16, Yq, and mainly the pericentromeric region of chromosome 9 was far more intensely labelled in amniocytes than in lymphocytes. Fluorescence in situ hybridisation with a specific classical satellite DNA probe, showed that this differential labelling was dependent on a greater sensitivity of chromosome 9 constitutive heterochromatin to exonuclease III digestion in amniocytes than in lymphocytes, thus indicating qualitative differences in this region between both human cellular materials. (J Med Genet 1995;32:32-35 In the present paper we present a modified in situ random primer extension procedure on exonuclease III treated chromosomes to show that there are exceptions to items (2) and (3) mentioned above. In fact, in the model we have tested, R bands coexist with labelled constitutive heterochromatin in chromosomes 1, 9, 16, and Yq. Moreover, this is shown to be specific to human amniocytes, human lymphocytes producing a different banding pattern. Materials and methods CULTURES AND METAPHASE PREPARATIONHuman amniotic fluid cells from four persons were obtained by amniocentesis, performed at 14 to 16 weeks' gestation, and cultured at 37°C in the dark with lOml Ham's F10 medium supplemented with 10% fetal calf serum, antibiotics, L-glutamine, hepes-buffer pH 7-2, and ultroser G. After a week of incubation, the amniocytes were subcultured, and harvested two days later by the trypsin suspension method or mechanically detached.Human peripheral blood lymphocytes were obtained from four normal persons, ranging from newborn to 35 years old. Cultures were set up for 72 hours at 37°C with 4 ml TC 199 medium supplemented with 10% fetal calf serum, antibiotics, and phytohaemagglutinin.Both amniocytes and lymphocytes were arrested at metaphase for three hours with 0 5 ztg/ ml colchicine, treated for 10 minutes with 0 075 mol/l KCI at 37°C, fixed three times in methanol:acetic acid (3:1), and spread on opposite sides of the same glass slide by the air dry method.

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“…Restriction endonucleases (RE-s) are known to induce DNA cleavage even in the fixed metaphase chromosomes (SAHASRABUDDHE et al 1978) and cause extraction of fragmented DNA (BIANCHI et al 1985), which can be revealed as characteristic staining patterns by Giemsa staining. Heterochromatic regions may show uniform or differentiated specific resistance or increased sensitivity to a RE (BABU and VERMA 1990;LOPEZ-FERNANDEZ et al 1991). Therefore, this method provides a powerful tool for the study of polymorphisms and compositional heterogeneity of the heterochromatic regions.…”

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Heterochromatin Heterogeneity Revealed by Restriction Endonuclease Digestion and Subsequent C-banding on Bovine Metaphase Chromosomes

Hidas¹

2004

Hereditas

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Restriction endonucleases in the study of eukaryotic chromosomes

Cited by 39 publications

References 92 publications

Karyological Comparison of Sphenops sepsoides, Chalcides chalcides, and C. ocellatus (Reptilia: Scincidae): Taxonomic Implications

Karyological Comparison of Sphenops sepsoides, Chalcides chalcides, and C. ocellatus (Reptilia: Scincidae): Taxonomic Implications

Difference in constitutive heterochromatin behaviour between human amniocytes and lymphocytes detected by a sequential in situ exonuclease III digestion-random primer extension procedure.

Heterochromatin Heterogeneity Revealed by Restriction Endonuclease Digestion and Subsequent C-banding on Bovine Metaphase Chromosomes

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