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Home / Equine Research Bank / Cytogenet Genome Res / An ordered BAC contig map of the equine major histocompatibi...
Cytogenetic and genome research2004; 102(1-4); 189-195; doi: 10.1159/000075747

An ordered BAC contig map of the equine major histocompatibility complex.

Abstract: A physical map of ordered bacterial artificial chromosome (BAC) clones was constructed to determine the genetic organization of the horse major histocompatibility complex. Human, cattle, pig, mouse, and rat MHC gene sequences were compared to identify highly conserved regions which served as source templates for the design of overgo primers. Thirty-five overgo probes were designed from 24 genes and used for hybridization screening of the equine USDA CHORI 241 BAC library. Two hundred thirty-eight BAC clones were assembled into two contigs spanning the horse MHC region. The first contig contains the MHC class II region and was reduced to a minimum tiling path of nine BAC clones that span approximately 800 kb and contain at least 20 genes. A minimum tiling path of a second contig containing the class III/I region is comprised of 14 BAC clones that span approximately 1.6 Mb and contain at least 34 genes. Fluorescence in situ hybridization (FISH) using representative clones from each of the three regions of the MHC localized the contigs onto ECA20q21 and oriented the regions relative to one another and the centromere. Dual-colored FISH revealed that the class I region is proximal to the centromere, the class II region is distal, and the class III region is located between class I and II. These data indicate that the equine MHC is a single gene-dense region similar in structure and organization to the human MHC and is not disrupted as in ruminants and pigs.
Copyright 2003 S. Karger AG, Basel
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Publication Date: 2004-02-19 PubMed ID: 14970701DOI: 10.1159/000075747Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't
  • Research Support
  • U.S. Gov't
  • Non-P.H.S.
  • Research Support
  • U.S. Gov't
  • P.H.S.

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research focuses on creating a physical map using ordered Bacterial Artificial Chromosome (BAC) clones to understand the genetic organization of the horse’s major histocompatibility complex (MHC). The map shows that the equine MHC is similarly structured and organized as the human MHC.

Construction of BAC Clone Map

  • The researchers created a physical map using ordered BAC clones to understand the genetic organization of the horse’s major histocompatibility complex (MHC).
  • This was done by comparing MHC gene sequences from humans, cattle, pigs, mice, and rats to identify regions that are highly conserved across these species.
  • These identified conserved regions were then utilized as source templates to design overgo primers – a key tool in genomics to probe specific sequences in the DNA.

Hybridization Screening

  • The researchers designed 35 overgo probes from 24 genes for hybridization screening of the equine USDA CHORI 241 BAC library.
  • This screening process led to the assembly of 238 BAC clones into two contigs, or overlapping sets of DNA, spanned across the horse MHC region.

Navigating the MHC Region

  • The first contig represents the MHC class II region and contains, at least, 20 genes. A minimum tiling path of nine BAC clones was used to span approximately 800 kilobases(Kb).
  • The second contig represents the class III/I region spanned by a tiling path of 14 BAC clones, covering approximately 1.6 megabases (Mb) and containing at least 34 genes.

Fluorescence In Situ Hybridization (FISH)

  • The researchers applied fluorescence in situ hybridization (FISH) using representative clones from each of the three regions of the MHC.
  • This technique allowed the researchers to accurately localize and orient the contigs onto ECA20q21 – a certain location on the horse’s chromosome.
  • The dual-colored FISH technique provided information about the location of the three MHC regions relative to the centromere – Class I region is closer to the centromere, Class II is farther, and Class III is located between Class I and II.

Conclusion

  • The findings from this research indicates that the equine MHC is a single gene-dense region similar in structure and organization to the human MHC.
  • This contrasts with the MHC of ruminants and pigs, which is disrupted.

Cite This Article

APA
Gustafson AL, Tallmadge RL, Ramlachan N, Miller D, Bird H, Antczak DF, Raudsepp T, Chowdhary BP, Skow LC. (2004). An ordered BAC contig map of the equine major histocompatibility complex. Cytogenet Genome Res, 102(1-4), 189-195. https://doi.org/10.1159/000075747

Publication

Cytogenetic and genome research
ISSN: 1424-859X
NlmUniqueID: 101142708
Country: Switzerland
Language: English
Volume: 102
Issue: 1-4
Pages: 189-195

Researcher Affiliations

Gustafson, A L
  • Department of Veterinary Anatomy and Public Health, College of Veterinary Medicine, Texas A&M University, College Station, TX 77843-4458, USA.
Tallmadge, R L
    Ramlachan, N
      Miller, D
        Bird, H
          Antczak, D F
            Raudsepp, T
              Chowdhary, B P
                Skow, L C

                  MeSH Terms

                  • Animals
                  • Blotting, Southern / methods
                  • Blotting, Southern / veterinary
                  • Chromosomes, Artificial, Bacterial / genetics
                  • Contig Mapping / methods
                  • Contig Mapping / veterinary
                  • DNA / genetics
                  • DNA Fingerprinting / methods
                  • DNA Fingerprinting / veterinary
                  • Horses / genetics
                  • In Situ Hybridization, Fluorescence / veterinary
                  • Major Histocompatibility Complex / genetics

                  Grant Funding

                  • HD-34086 / NICHD NIH HHS

                  Citations

                  This article has been cited 30 times.
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