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Related Experiment Videos

Cloning and expression of caprine interferon-gamma

J C Beyer1, R W Stich, D S Hoover

  • 1Department of Veterinary Microbiology, Pathology, Washington State University, Pullman, WA 99164-7040, USA. jcb@vetmed.wsu.edu

Gene
|May 23, 1998
PubMed
Summary

Researchers successfully cloned and expressed caprine interferon-gamma (IFN-gamma) cDNA. The recombinant protein demonstrated biological activity and conserved antigenic epitopes, paving the way for further research into goat immune responses.

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Area of Science:

  • Immunology
  • Molecular Biology
  • Biotechnology

Background:

  • Interferon-gamma (IFN-gamma) is a crucial cytokine involved in immune responses.
  • Cloning and expression of caprine IFN-gamma are essential for studying its function in goats.

Purpose of the Study:

  • To clone and characterize caprine IFN-gamma cDNA.
  • To express recombinant caprine IFN-gamma (rCaIFN-gamma) in vitro and in vivo.
  • To assess the biological activity and antigenic properties of rCaIFN-gamma.

Main Methods:

  • Reverse transcription-polymerase chain reaction (RT-PCR) for cDNA cloning.
  • Subcloning into expression plasmids (CMV promoter and CAEV LTR).
  • Transfection of COS-7 cells and expression analysis.
  • Vesicular stomatitis virus (VSV) cytopathic effect reduction assay for biological activity.

Related Experiment Videos

  • Intramuscular injection in mice and Western blot for in vivo expression confirmation.

    • Main Results:

    • Caprine IFN-gamma cDNA (498bp) was successfully cloned, encoding a 166 amino acid protein.
    • High sequence homology was observed between caprine IFN-gamma and related species (ovine, bovine, cervine).
    • Recombinant caprine IFN-gamma exhibited biological activity and shared antigenic epitopes with bovine IFN-gamma.
    • In vivo expression was confirmed using the CAEV LTR in mice.

    Conclusions:

    • The successful cloning and expression of functional caprine IFN-gamma provide a valuable tool for immunological studies in goats.
    • The high sequence homology suggests conserved functions across ruminant species.
    • This work facilitates further investigation into the role of IFN-gamma in caprine health and disease.