Molecular cloning of an equine satellite-type DNA sequence and its chromosomal localization.
Abstract: We have molecularly cloned portions of equine satellite-type DNA and investigated the organization of the DNA sequence of the cloned segments. Sequence analysis and dot-blot analysis, using the cloned sequence (ES200) as a probe, indicate that the satellite-type DNA sequence consists mainly of 221-bp tandem repeats and represents 3.7-11% of the equine genome. Southern blot analysis further shows that (1) no sequences homologous to ES200 exist in the human, swine, and bovine genomes and that (2) the fragment pattern of the satellite-type DNA produced by ApaI cleavage shows a slight difference among equine DNA samples, indicating a slight sequence variation between individual horses. To localize the satellite-type DNA in the horse genome, chromosome spreads on glass slides were first subjected to G-banding to identify each chromosome and then hybridized in situ with biotin-labeled ES200. The chromosome banding, together with the hybridization, demonstrates that the satellite sequences are preferentially localized on the centromeric regions of all chromosomes except chromosomes 2, 9, and 11.
Publication Date: 1994-01-01 PubMed ID: 8275703DOI: 10.1159/000133657Google Scholar: Lookup
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- Comparative Study
- Journal Article
Summary
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This research involved the molecular cloning of portions of horse DNA of a particular type known as satellite DNA. Findings indicate that this type of DNA consists mainly of repeated sequences and represents a significant proportion of the horse’s genome. It was also discovered that there are slight differences in this sequencing among individual horses.
Study Details and Methods
- The study focused on performing molecular cloning of equine satellite-type DNA, a form of highly repetitive DNA mainly found on the chromosome’s centromeric and pericentromeric regions.
- The cloned segments were then investigated for their DNA sequence organization with sequence analysis and dot-blot analysis used as testing methods. A cloned sequence called ES200 was used as a probe in this respect.
- The research also involved Southern blot analysis which confirmed that no sequences similar to ES200 were found in the human, swine, and bovine genomes.
- The Southern blot analysis also displayed the segment pattern of the satellite-type DNA produced by ApaI cleavage. This revealed slight variations in sequence among different horse DNA samples.
Results and Key Findings
- The study found out that the satellite-type DNA sequence predominantly comprises of 221-bp tandem repeats and makes up between 3.7% and 11% of the equine genome.
- The investigation showed that there are slight differences in the sequence of satellite DNA among individual horses’ DNA samples.
- To define the location of the satellite-type DNA in the horse genome, the research implemented chromosome spreads on glass slides. They first applied G-banding on the slides to recognize each chromosome and then used biotin-labeled ES200 for in-situ hybridization.
- This process showed that the satellite sequences are primarily found in the centromeric regions of all chromosomes except for chromosomes 2, 9 and 11. This means the centromeric region of most equine chromosomes is rich in satellite DNA.
Conclusions
- The study revealed significant insights into the organization of satellite DNA in equines, its quantity in the genome, and its chromosomal localization.
- These findings contribute to our understanding of the genetic make-up of horses and may help in making comparisons between species, understanding individual genetic variations among horses, and studying the functions of satellite DNA and its role in genomic stability and chromosomal behavior.
Cite This Article
APA
Sakagami M, Hirota K, Awata T, Yasue H.
(1994).
Molecular cloning of an equine satellite-type DNA sequence and its chromosomal localization.
Cytogenet Cell Genet, 66(1), 27-30.
https://doi.org/10.1159/000133657 Publication
Researcher Affiliations
- Laboratory of Molecular and Cellular Biology, Equine Research Institute, Japan Racing Association, Ibaraki.
MeSH Terms
- Animals
- Base Sequence
- Blotting, Southern
- Cattle
- Chromosome Banding
- Chromosome Mapping
- DNA, Satellite / analysis
- DNA, Satellite / genetics
- Horses / genetics
- Humans
- Molecular Sequence Data
- Sequence Homology, Nucleic Acid
- Species Specificity
- Swine
Citations
This article has been cited 2 times.- Piras FM, Cappelletti E, Abdelgadir WA, Salamon G, Vignati S, Santagostino M, Sola L, Nergadze SG, Giulotto E. A Satellite-Free Centromere in Equus przewalskii Chromosome 10.. Int J Mol Sci 2023 Feb 18;24(4).
- Piras FM, Nergadze SG, Magnani E, Bertoni L, Attolini C, Khoriauli L, Raimondi E, Giulotto E. Uncoupling of satellite DNA and centromeric function in the genus Equus.. PLoS Genet 2010 Feb 12;6(2):e1000845.
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