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Viral sequences are associated with many histocompatibility genes.
Immunogenetics
|January 1, 1986
Summary
Researchers cloned a retroviral gene fragment and identified 31 restriction fragment length polymorphisms (RFLPs). These RFLPs link viral sequences to 15 minor histocompatibility (H) loci, aiding in gene mapping and cloning efforts.
Area of Science:
- Genetics
- Molecular Biology
- Immunology
Background:
- Murine retroviruses possess genes crucial for viral integration, such as the endonuclease domain within the pol gene.
- Minor histocompatibility (H) loci are important in immune responses and transplantation, but their genetic basis is often poorly understood.
- Restriction Fragment Length Polymorphisms (RFLPs) are valuable tools for genetic mapping and identifying disease-associated genes.
Purpose of the Study:
- To clone and characterize a specific murine retroviral gene fragment.
- To identify and map restriction fragment length polymorphisms (RFLPs) associated with this fragment.
- To investigate the potential linkage between viral gene sequences and minor histocompatibility (H) loci for gene mapping.
Main Methods:
- Cloning of a 5.5 kb Pvu II polymorphic restriction fragment from C57BL/6By mice using a spleen focus-forming env probe.
- Subcloning of a 358 bp fragment from the cloned DNA.
- Hybridization and sequencing studies to identify the retroviral gene region encoding endonuclease.
- Analysis of murine genomic DNAs using Southern blotting with the 358 bp probe to detect RFLPs.
- Mapping of identified RFLPs to known minor histocompatibility (H) loci.
Main Results:
- A 358 bp fragment encoding a murine retroviral endonuclease critical for integration was cloned and subcloned.
- Hybridization of this probe to digested murine genomic DNA revealed 31 distinct RFLPs.
- Sixteen of these RFLPs were localized near 15 specific minor histocompatibility (H) loci (H-3, H-4, H-7, H-13, H-15, H-16, H-17, H-19, H-22, H-24, H-27, H-30, H-34, H-36, H-38).
- Tentative assignment of H-17, H-34, and H-38 to chromosome 12 was made.
- Observation that H-2 congenic strains retain chromosome 12 segments from parental strains.
Conclusions:
- The proximity of cloned viral sequences to multiple minor H loci suggests evolutionary and functional significance.
- The identified RFLPs provide a valuable resource for facilitating the cloning of minor H genes.
- The study contributes to the genetic mapping of minor histocompatibility loci and chromosome 12.