Localisation of satellite DNA sequences on human metaphase chromosomes using bromodeoxyuridine-labelled probes

Chromosoma ◽  
1988 ◽  
Vol 97 (1) ◽  
pp. 11-18 ◽  
Author(s):  
M. Frommer ◽  
C. Paul ◽  
P. C. Vincent
Keyword(s):  
Dna Sequences ◽  
Satellite Dna ◽  
Metaphase Chromosomes

scholarly journals Satellite DNA induces unstable expression of the adjacent herpes simplex virus tk gene cotransfected in mouse cells.

1988 ◽  
Vol 8 (3) ◽  
pp. 1336-1344 ◽  
Author(s):  
D Talarico ◽  
A F Peverali ◽  
E Ginelli ◽  
R Meneveri ◽  
C Mondello ◽  
...  
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Dna Sequences ◽  
Satellite Dna ◽  
Repetitive Sequences ◽  
Metaphase Chromosomes ◽  
Highly Repetitive Dna ◽  
Cellular Dna ◽  
Mouse Satellite Dna ◽  
Simplex Virus

To study the influence of clustered highly repetitive DNA sequences on the expression of adjacent genes, LTK- cells were cotransfected with the herpes simplex virus thymidine kinase (tk) gene and mouse satellite DNA. TK+ transformants containing a few copies of the tk genes flanked by satellite DNA were isolated. In situ hybridization on the metaphase chromosomes indicated that in each cell line the TK sequences resided at a single chromosomal site and that integration occurred preferentially into regions of the cellular DNA rich in highly repetitive sequences. The prominent feature of these cell lines was their phenotypic instability. Suppression and reexpression of the tk gene occurred at high frequency (greater than 3%) and did not correlate with any significant change in the organization of foreign DNA or with the presence of selective agents. These results indicate that satellite DNA, the major component of constitutive heterochromatin, may influence the expression of adjacent genes by affecting the chromatin structure.

Satellite DNA induces unstable expression of the adjacent herpes simplex virus tk gene cotransfected in mouse cells

1988 ◽  
Vol 8 (3) ◽  
pp. 1336-1344
Author(s):  
D Talarico ◽  
A F Peverali ◽  
E Ginelli ◽  
R Meneveri ◽  
C Mondello ◽  
...  
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Dna Sequences ◽  
Satellite Dna ◽  
Repetitive Sequences ◽  
Metaphase Chromosomes ◽  
Highly Repetitive Dna ◽  
Cellular Dna ◽  
Mouse Satellite Dna ◽  
Simplex Virus

To study the influence of clustered highly repetitive DNA sequences on the expression of adjacent genes, LTK- cells were cotransfected with the herpes simplex virus thymidine kinase (tk) gene and mouse satellite DNA. TK+ transformants containing a few copies of the tk genes flanked by satellite DNA were isolated. In situ hybridization on the metaphase chromosomes indicated that in each cell line the TK sequences resided at a single chromosomal site and that integration occurred preferentially into regions of the cellular DNA rich in highly repetitive sequences. The prominent feature of these cell lines was their phenotypic instability. Suppression and reexpression of the tk gene occurred at high frequency (greater than 3%) and did not correlate with any significant change in the organization of foreign DNA or with the presence of selective agents. These results indicate that satellite DNA, the major component of constitutive heterochromatin, may influence the expression of adjacent genes by affecting the chromatin structure.

DNA sequence mapping in interphase and metaphase chromosomes by fluorescence in situ hybridization

1992 ◽  
Vol 50 (1) ◽  
pp. 496-497
Author(s):  
Barbara Trask ◽  
Susan Allen ◽  
Anne Bergmann ◽  
Mari Christensen ◽  
Anne Fertitta ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Dna Sequence ◽  
Dna Sequences ◽  
Dual Band ◽  
Nick Translation ◽  
Metaphase Chromosomes ◽  
Band Pass ◽  
Texas Red ◽  
Fluorescent Spot

Using fluorescence in situ hybridization (FISH), the positions of DNA sequences can be discretely marked with a fluorescent spot. The efficiency of marking DNA sequences of the size cloned in cosmids is 90-95%, and the fluorescent spots produced after FISH are ≈0.3 μm in diameter. Sites of two sequences can be distinguished using two-color FISH. Different reporter molecules, such as biotin or digoxigenin, are incorporated into DNA sequence probes by nick translation. These reporter molecules are labeled after hybridization with different fluorochromes, e.g., FITC and Texas Red. The development of dual band pass filters (Chromatechnology) allows these fluorochromes to be photographed simultaneously without registration shift.

scholarly journals Transcription termination and processing of transcripts from tRNA-related Xenopus satellite DNA sequences

1987 ◽  
Vol 164 (2) ◽  
pp. 287-293 ◽  
Author(s):  
Wolfgang MEYERHOF ◽  
Burghardt WITTIG ◽  
Beatrix TAPPESER ◽  
Walter KNOCHEL
Keyword(s):  
Dna Sequences ◽  
Satellite Dna ◽  
Transcription Termination

scholarly journals Repetitive DNA sequences accelerate molecular cytogenetic research in plants with small chromosomes

2019 ◽  
Vol 24 (2) ◽  
pp. 82
Author(s):  
Agus Budi Setiawan ◽  
Ari Wibowo ◽  
Chee How Teo ◽  
Shinji Kikuchi ◽  
Takato Koba
Keyword(s):  
Repetitive Dna ◽  
Dna Sequences ◽  
5S Rdna ◽  
Repetitive Dna Sequences ◽  
Type I ◽  
Chromosome Identification ◽  
Metaphase Chromosomes ◽  
Homologous Chromosomes ◽  
Centromeric Sequence ◽  
Subtelomeric Sequence

Repetitive DNA sequences are highly abundant in plant genomes and are favorable probes for chromosome identification in plants. However, it is difficult to conduct studies on the details of metaphase chromosome structures in plants with small chromosomes due to their highly condensed status. Therefore, identification of homologous chromosomes for karyotyping and analyzing chromosome structures is a challenging issue for cytogeneticists without specific probes and precise chromosome stages. In this study, five repetitive DNA probes, i.e., 5S and 45S ribosomal DNAs (rDNAs), melon centromeric sequence (Cmcent), cucumber subtelomeric sequence (Type I), and microsatellite (CT)10 repeats, were used to identify primary constrictions and homologous chromosomes for karyotyping. Four and two loci of 45S rDNA were respectively observed on metaphase and pachytene chromosomes of Abelia × grandiflora. Cmcent was detected on both primary constrictions of melon pachytene and metaphase chromosomes. Furthermore, one pair of 5S rDNA signals were hybridized on melon metaphase chromosomes. Eight and two loci of 45S and 5S rDNA were respectively detected on cucumber chromosomes. Type I and (CT)10 probes were specifically hybridized on subtelomeric and interstitial regions on the chromosomes, respectively. These results suggest that repetitive DNA sequences are versatile probes for chromosome identification in plants with small chromosomes, particularly for karyotyping analyses.

Epigenetic control of mammalian centromere protein binding: does DNA methylation have a role?

1996 ◽  
Vol 109 (9) ◽  
pp. 2199-2206
Author(s):  
A.R. Mitchell ◽  
P. Jeppesen ◽  
L. Nicol ◽  
H. Morrison ◽  
D. Kipling
Keyword(s):  
Dna Sequences ◽  
Satellite Dna ◽  
Inbred Mouse ◽  
Inbred Mouse Strain ◽  
Chromosome 1 ◽  
Satellite Sequences ◽  
E Protein ◽  
Minor Satellite ◽  
Internal Block ◽  
Terminal Block

Chromosome 1 of the inbred mouse strain DBA/2 has a polymorphism associated with the minor satellite DNA at its centromere. The more terminal block of satellite DNA sequences on this chromosome acts as the centromere as shown by the binding of CREST ACA serum, anti-CENP-B and anti-CENP-E polyclonal sera. Demethylation of the minor satellite DNA sequences accomplished by growing cells in the presence of the drug 5-aza-2′-deoxycytidine results in a redistribution of the CENP-B protein. This protein now binds to an enlarged area on the more terminal block and in addition it now binds to the more internal block of minor satellite DNA sequences on chromosome 1. The binding of the CENP-E protein does not appear to be affected by demethylation of the minor satellite sequences. We present a model to explain these observations. This model may also indicate the mechanism by which the CENP-B protein recognises specific sites within the arrays of minor satellite DNA on mouse chromosomes.

Chromatin organization in the homogeneously staining regions of a methotrexate-resistant mouse cell line: interspersion of inactive and active chromatin domains distinguished by acetylation of histone H4

1996 ◽  
Vol 109 (9) ◽  
pp. 2221-2228 ◽  
Author(s):  
L. Nicol ◽  
P. Jeppesen
Keyword(s):  
Cell Line ◽  
Dna Sequences ◽  
Satellite Dna ◽  
Mouse Cell ◽  
Histone H4 ◽  
Active Chromatin ◽  
Chromatin Domains ◽  
Resistant Mouse ◽  
Major Satellite ◽  
Drosophila Heterochromatin

We have analyzed the organization of the homogeneously staining regions (HSRs) in chromosomes from a methotrexate-resistant mouse melanoma cell line. Fluorescence in situ hybridization techniques were used to localize satellite DNA sequences and the amplified copies of the dihydrofolate reductase (DHFR) gene that confer drug-resistance, in combination with immunofluorescence using antibody probes to differentiate chromatin structure. We show that the major DNA species contained in the HSRs is mouse major satellite, confirming previous reports, and that this is interspersed with DHFR DNA in an alternating tandem array that can be resolved at the cytological level. Mouse minor satellite DNA, which is normally located at centromeres, is also distributed along the HSRs, but does not appear to interfere with centromere function. The blocks of major satellite DNA are coincident with chromatin domains that are labelled by an autoantibody that recognizes a mammalian homologue of Drosophila heterochromatin-associated protein 1, shown previously to be confined to centric heterochromatin in mouse. An antiserum that specifically recognizes acetylated histone H4, a marker for active chromatin, fails to bind to the satellite DNA domains, but labels the intervening segments containing DHFR DNA. We can find no evidence for the spreading of the inactive chromatin domains into adjacent active chromatin, even after extended passaging of cells in the absence of methotrexate selection.

261 LABELING SEX-SPECIFIC DNA SEQUENCES IN MAMMALIAN SPERM

2008 ◽  
Vol 20 (1) ◽  
pp. 210
Author(s):  
B. A. Didion ◽  
R. Bleher
Keyword(s):  
Dna Sequences ◽  
Somatic Cells ◽  
Complete Separation ◽  
Cell Nuclei ◽  
Metaphase Chromosomes ◽  
Telomeric Sequences ◽  
Synthetic Dna ◽  
Y Chromosomes ◽  
Boar Sperm ◽  
The Difference

While flow cytometric separation of X- andY-chromosome- bearing sperm has advanced to the point of acceptance in the commercial production of sex-preselected cattle, it is important to continue researching this area to improve efficiencies. For example, the difference in DNA sequence between the X- andY-chromosomes has merit as a foundation for an alternative sperm sexing approach that could enable the complete separation and use of an entire ejaculate. We used synthetic DNA mimics conjugated to a fluorescent dye for in situ detection of Y-chromosomes in metaphase preparations of porcine somatic cells and spermatozoa. Peptide nucleic acids (PNA) are synthetic compounds with higher affinity and stability than conventional DNA probes and are used as specific hybridization probes to complementary DNA. The application of PNA probes was demonstrated previously in telomere analysis studies, and we confirmed their efficacy using a CY3-(CCCTAA)3 PNA to probe bull and boar sperm telomeric sequences. Using male porcine somatic cells and theY-chromosome as a template, we arranged for the synthesis of a CY3-conjugated PNA to bind 13-15 base pairs of unique, Y-chromosome sequence. By testing different labeling conditions, we found that brief incubation of metaphase chromosomes with the PNA produced a localized signal on theY-chromosome. No signals were present when chromosomes of porcine female somatic cells were incubated with the PNA probes. Because chromosomes occupy non-random territories in all cell nuclei including those in sperm, we expected to find centrally located signals in 50% of fixed boar sperm when these were treated with the same PNA as used for the somatic cells. We found the signals present in 161 of 302 (53.3%) sperm to consist of a single, centrally located, round fluorescent dot in the sperm head. Further research is required to establish the uptake of PNA in live sperm toward evaluation of this approach for sperm sexing.

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