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Engineering the transmissible gastroenteritis virus genome as an expression vector inducing lactogenic immunity
Isabel Sola1, Sara Alonso, Sonia Zúñiga
1Centro Nacional de Biotecnología, CSIC, Department of Molecular and Cell Biology, Campus Universidad Autónoma, Cantoblanco, Madrid, Spain.
Journal of Virology
|March 14, 2003
Summary
Transmissible gastroenteritis virus (TGEV) was engineered into an expression vector for stable heterologous gene expression. This TGEV vector shows promise for vaccine development and gene therapy applications.
Area of Science:
- Virology
- Molecular Biology
- Gene Expression
Background:
- Transmissible gastroenteritis virus (TGEV) is a significant swine pathogen.
- Engineering viral genomes offers potential for novel biotechnological tools.
Purpose of the Study:
- To engineer the TGEV genome as an expression vector for stable heterologous gene expression.
- To evaluate the efficacy of TGEV-based expression systems and their potential applications.
Main Methods:
- Infectious cDNA of TGEV was engineered to express a heterologous gene (GFP).
- Transcription-regulating sequences (TRS) from ORF 3a were utilized for gene expression.
- Recombinant viruses were assessed for gene expression stability, pathogenicity, and immunogenicity.
Main Results:
- Efficient and stable expression of GFP (>40 μg/10^6 cells, >20 passages) was achieved using TGEV ORF 3a TRS.
- Recombinant TGEV retained enteropathogenicity with reduced enteric tissue growth.
- A specific lactogenic immune response was elicited against the heterologous protein in sows and progeny.
- Alternative insertion sites led to instability and altered 3' end genetic organization.
Conclusions:
- The engineered TGEV vector provides high-level, stable gene expression comparable to established vectors.
- This TGEV-based system holds significant promise for developing vaccines and potential gene therapy strategies.
- Understanding TRS function is crucial for optimizing recombinant viral vector design.