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Related Experiment Videos

Design and construction of African swine fever virus chimeras incorporating foreign viral epitopes.

A Brun1, F Rodríguez, F Parra

  • 1Centro de Investigación en Sanidad Animal (CISA-INIA), Valdeolmos, Madrid, Spain.

Archives of Virology
|September 11, 1999
PubMed
Summary

Researchers genetically modified African swine fever virus (ASFV) to insert foreign epitopes into its structure. This study demonstrates the feasibility of incorporating foreign sequences into ASFV particles without impacting virus viability.

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Area of Science:

  • Virology
  • Molecular Biology
  • Genetic Engineering

Background:

  • African swine fever virus (ASFV) is a significant pathogen affecting swine populations globally.
  • Developing novel strategies for studying and potentially controlling ASFV is crucial.
  • Genetic manipulation of viral particles offers a platform for introducing foreign antigens.

Purpose of the Study:

  • To investigate the feasibility of introducing foreign epitopes into the ASFV particle via genetic manipulation.
  • To develop and test transfer vectors for inserting foreign DNA sequences into ASFV.
  • To assess the viability and expression of modified ASFV particles.

Main Methods:

  • Developed transfer vectors containing the ASFV p54 protein gene with inserted foreign epitope sequences (FMDV VP1 site A, TGEV N DA3 determinant).

Related Experiment Videos

  • Inserted chimeric p54 genes into the ASFV genome using homologous recombination at the thymidine kinase (TK) locus.
  • Analyzed recombinant viruses for expression of chimeric proteins, recognition by antibodies, presence in viral particles, and replication in Vero cells.

    • Main Results:

    • Recombinant ASFV efficiently expressed chimeric p54 proteins containing foreign epitopes.
    • Chimeric proteins were recognized by antibodies specific to both ASFV p54 and the foreign epitopes.
    • Modified p54 proteins were incorporated into viral particles and functionally complemented wild-type p54, supporting virus replication.
    • Deletion of the p54 gene in recombinant viruses did not affect replication in Vero cells.

    Conclusions:

    • Demonstrated the feasibility of incorporating foreign amino acid sequences (up to 18 residues) into an ASFV protein component.
    • Showcased that genetic modification of ASFV can yield viable viral particles expressing foreign epitopes.
    • This approach provides a foundation for developing novel ASFV-based tools for research or potential vaccine strategies.