Cloning and characterization of the equine F18 gene, which has a novel exon.
Abstract: A genomic clone isolated from an equine genomic library probed with an oligonucleotide (CAG)10 showed high sequence similarity to the human F18 gene and was tentatively named equine F18 gene. Because the human F18 gene is expressed in many tissues, we examined whether this equine clone was also expressed in equine tissues. The cDNA encoding equine F18 was obtained by the reverse transcriptase-polymerase chain reaction (RT-PCR) from equine thymus. The nucleotide sequence of the equine F18 cDNA (1940 bp) was determined and contained both the ATG initiation codon and a poly(A) sequence. The cDNA sequence contained sequence homologous to exon 3 of human F18 gene and a new exon that is not found in the human F18 gene. The equine F18 gene contains a CAG repetitive sequence (CRS) that is translated into a polyglutamine tract. The CRS was analyzed for polymorphism by PCR: four and two different alleles were observed with unrelated east-Asian and thoroughbred horses, respectively. The equine F18 gene was mapped to Xq29.1 by fluorescence in situ hybridization. The human F18 gene is also X-linked. These data strongly supported the conclusion that the clone contains the equine homologue of the human F18 gene.
Publication Date: 1998-11-04 PubMed ID: 9800327DOI: 10.1046/j.1365-2052.1998.295356.xGoogle Scholar: Lookup
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- Comparative Study
- Journal Article
Summary
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The research article discusses the cloning and characterization of the equine F18 gene from horses, which was found to have a high sequence similarity to the human F18 gene and contained a new exon not found in the human version.
Identifying the Equine F18 Gene
- This study began with the isolation of a genomic clone from an equine genomic library. This clone was identified by probing the library with an oligonucleotide (CAG)10.
- Upon examination of this clone, the researchers found that it displayed a high degree of sequence similarity to the human F18 gene, leading them to tentatively name this clone the equine F18 gene.
Expression of the Equine F18 Gene
- Given that the human F18 gene is expressed in a multitude of tissues, the researchers sought to determine whether the equine F18 gene was likewise expressed in equine tissues.
- For this purpose, they acquired cDNA encoding equine F18 through a reverse transcriptase-polymerase chain reaction (RT-PCR) from equine thymus tissue.
- After determining the nucleotide sequence of this cDNA, they found it contained both the ATG initiation codon and a poly(A) sequence.
Discovery of a New Exon
- More interestingly, the cDNA sequence also presented sequence homology to exon 3 of the human F18 gene and an entirely new exon that is not found in the human F18 gene.
- This suggests that the equine F18 gene has developed its own unique exon that may be central to its distinctive functionality in equine biology.
Further Analysis and Mapping
- The equine F18 gene contains a CAG repetitive sequence (CRS) that is translated into a polyglutamine tract.
- The CRS was analyzed for polymorphism via PCR, revealing four different alleles in unrelated east-Asian horses and two different alleles in thoroughbred horses.
- The precise location of the equine F18 gene was determined as Xq29.1 through a process known as fluorescence in situ hybridization. This aligns with the human F18 gene’s location, which is also X-linked.
- The convergence of these results strongly supports the hypothesis that the clone contains the equine equivalent of the human F18 gene.
Cite This Article
APA
Tozaki T, Hirota K, Mashima S, Tomita M, Mukoyama H.
(1998).
Cloning and characterization of the equine F18 gene, which has a novel exon.
Anim Genet, 29(5), 381-384.
https://doi.org/10.1046/j.1365-2052.1998.295356.x Publication
Researcher Affiliations
- Department of Molecular Genetics, Laboratory of Racing Chemistry, Tochigiken, Japan.
MeSH Terms
- Animals
- Base Sequence
- Cloning, Molecular
- DNA Primers / genetics
- DNA, Complementary / genetics
- Exons
- Genetic Linkage
- Genomic Library
- Horses / genetics
- Humans
- In Situ Hybridization, Fluorescence
- Molecular Sequence Data
- Polymerase Chain Reaction
- Polymorphism, Genetic
- Proteins / genetics
- Species Specificity
- Trinucleotide Repeats
- X Chromosome / genetics
Citations
This article has been cited 1 times.- Shiue Y-L, Millon LV, Skow LC, Honeycutt D, Murray JD, Bowling AT. Synteny and regional marker order assignment of 26 type I and microsatellite markers to the horse X- and Y-chromosomes. Chromosome Res 2000;8(1):45-55.
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