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Porcine adenovirus-3 as a helper-dependent expression vector
P Seshidhar Reddy1, Neeraja Idamakanti1, Lorne A Babiuk1
1Virology Group, Veterinary Infectious Disease Organization, 120 Veterinary Road, University of Saskatchewan, Saskatoon, Saskatchewan , Canada S7N 5E31.
The Journal of General Virology
|December 2, 1999
Summary
Porcine adenovirus-3 (PAV-3) vectors were developed using novel complementing cell lines. These helper-dependent PAV-3 recombinants efficiently express GFP but do not replicate in non-porcine cells, suggesting vaccine vector potential.
Area of Science:
- Virology
- Molecular Biology
- Vaccine Development
Background:
- Porcine adenovirus-3 (PAV-3) is a candidate for novel pig vaccine vectors.
- Development of helper-dependent vectors requires complementing cell lines.
Purpose of the Study:
- To create E1-complementing porcine cell lines for helper-dependent PAV-3 vector generation.
- To construct and characterize a helper-dependent PAV-3 vector expressing GFP.
Main Methods:
- Generated porcine cell lines expressing human adenovirus-5 (HAV-5) E1 proteins.
- Constructed and propagated PAV-3 recombinants with E3 and E1A deletions.
- Inserted the GFP gene into the E1A region of a PAV-3 recombinant.
- Assessed viral entry and replication in various cell types.
Main Results:
- HAV-5 E1A proteins transformed porcine cells and complemented PAV-3 E1A.
- A helper-dependent PAV-3 vector (PAV202) expressing GFP was successfully constructed.
- PAV-3 enters but does not replicate in dog, sheep, bovine, and human cells.
Conclusions:
- Developed functional complementing cell lines for PAV-3 vector production.
- Demonstrated the potential of helper-dependent PAV-3 vectors for vaccine applications.
- PAV-3 shows species-specific replication, limiting its spread to non-target cells.