Analysis of ELA-DQB exon 2 polymorphism in Argentine Creole horses by PCR-RFLP and PCR-SSCP.
Abstract: The second exon of equine leucocyte antigen (ELA)-DQB genes was amplified from genomic DNA of 32 Argentine Creole horses by PCR. Amplified DNA was analysed by PCR-restriction fragment length polymorphism (RFLP) and PCR-single-strand conformation polymorphism (SSCP). The PCR-RFLP analysis revealed two HaeIII patterns, four RsaI patterns, five MspI patterns and two HinfI patterns. EcoRI showed no variation in the analysed sample. Additional patterns that did not account for known exon 2 DNA sequences were observed, suggesting the existence of novel ELA-DQB alleles. PCR-SSCP analysis exhibited seven different band patterns, and the number of bands per animal ranged from four to nine. Both methods indicated that at least two DQB genes are present. The presence of more than two alleles in each animal showed that the primers employed in this work are not specific for a unique DQB locus. The improvement of this PCR-RFLP method should provide a simple and rapid technique for an accurate definition of ELA-DQB typing in horses.
Publication Date: 2003-07-31 PubMed ID: 12887619DOI: 10.1046/j.1439-0442.2003.00543.xGoogle Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The study focuses on understanding the genetic diversity of Argentine Creole horses by examining variations in a specific gene. Using scientific procedures, the researchers found several different patterns of genetic data, suggesting that there might be previously unknown versions of this gene.
Objective of the Study
- The main objective of this study was to identify polymorphisms (variations) in the second exon (a section) of equine leucocyte antigen (ELA)-DQB genes in Argentine Creole horses. This sort of variation often plays a significant role in diversity within a species and could influence health, performance traits and susceptibility to diseases. Hence knowing this can be crucial.
Conducting the Research
- For the purpose of the study, DNA was extracted from 32 Argentine Creole horses and amplified using Polymerase Chain Reaction (PCR), a commonly used method in molecular biology to make multiple copies of a specific DNA segment.
- The amplified DNA was then analysed using two techniques: PCR- restriction fragment length polymorphism (RFLP) and PCR-single-strand conformation polymorphism (SSCP). These methods are used to detect variations in genetic sequences.
Findings
- The PCR-RFLP analysis revealed two HaeIII patterns, four RsaI patterns, five MspI patterns and two HinfI patterns. In simpler terms, these patterns represent different versions of the gene being examined. The variation between different horses was much higher than expected from known versions of the gene, suggesting that there may be new, undiscovered alleles (versions of the gene).
- The PCR-SSCP analysis exhibited seven different band patterns, and the number of bands (differentiate specific DNA product resulted from PCR) observed per animal ranged between four to nine.
- These findings indicated the existence of at least two DQB genes in these horses, which revealed a high level of genetic diversity within this breed. Also, the presence of more than two alleles in each animal indicated that the primers (short DNA pieces that are the starting point for DNA synthesis) used in this study are not specific for a single DQB location, revealing even more genetic complexity.
Implication of Findings
- The results of the study imply the existence of novel ELA-DQB alleles, enhancing the understanding of genetic diversity within the Argentine Creole horse population.
- The discovery could potentially lead to the development of more accurate techniques for gene typing of horses, which may influence breeding strategies and efforts towards the conservation of genetic diversity within the species.
- Lastly, study proposes improvements in the PCR-RFLP method for it to be a simple and rapid technique for an accurate definition of ELA-DQB typing in horses.
Cite This Article
APA
Villegas-Castagnasso EE, Díaz S, Giovambattista G, Dulout FN, Peral-García P.
(2003).
Analysis of ELA-DQB exon 2 polymorphism in Argentine Creole horses by PCR-RFLP and PCR-SSCP.
J Vet Med A Physiol Pathol Clin Med, 50(6), 280-285.
https://doi.org/10.1046/j.1439-0442.2003.00543.x Publication
Researcher Affiliations
- Centro de Investigaciones en Genética Básica y Aplicada, Facultad de Ciencias Veterinarias, Universidad Nacional de La Plata, 60 y 118 C.C. 296, C.P. B1900AVW, La Plata, Argentina.
MeSH Terms
- Animals
- Argentina
- Breeding
- DNA Primers
- Exons / genetics
- Genetic Variation
- Histocompatibility Antigens Class II / classification
- Histocompatibility Antigens Class II / genetics
- Horses / blood
- Horses / genetics
- Horses / immunology
- Phylogeny
- Polymerase Chain Reaction / veterinary
- Polymorphism, Genetic
- Polymorphism, Restriction Fragment Length
- Polymorphism, Single-Stranded Conformational
- Restriction Mapping / veterinary
Citations
This article has been cited 3 times.- Klumplerova M, Splichalova P, Oppelt J, Futas J, Kohutova A, Musilova P, Kubickova S, Vodicka R, Orlando L, Horin P. Genetic diversity, evolution and selection in the major histocompatibility complex DRB and DQB loci in the family Equidae.. BMC Genomics 2020 Sep 30;21(1):677.
- Miller D, Tallmadge RL, Binns M, Zhu B, Mohamoud YA, Ahmed A, Brooks SA, Antczak DF. Polymorphism at expressed DQ and DR loci in five common equine MHC haplotypes.. Immunogenetics 2017 Mar;69(3):145-156.
- Ross P, Buntzman AS, Vincent BG, Grover EN, Gojanovich GS, Collins EJ, Frelinger JA, Hess PR. Allelic diversity at the DLA-88 locus in Golden Retriever and Boxer breeds is limited.. Tissue Antigens 2012 Aug;80(2):175-83.
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