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OJVRTM
Online Journal of Veterinary Research©
Volume 9 (1): 57-65, 2005
PCR-based plasmid vector for recombinant bovine herpesvirus
Souza VF, Dellagostin OA , Roehe PM.
Centre of Veterinary
Research Desidério Finamor, Centre of Biotechnology, Federal University of
Pelotas, Department
of Microbiology, Virology Laboratory, Federal University of Rio Grande do Sul, RS, Brazil.
ABSTRACT
Souza VF, Dellagostin OA, Roehe PM. PCR-based
plasmid vector for recombinant bovine herpesvirus, Online J Vet Res 9 (1): 57-65, 2005. A PCR-based plasmid vector for
recombinant Bovine Herpesvirus type 5 (BHV-5) is described. The protocol
simplifies viral DNA restriction endonucleases digestion and cloning methods. PCR was
used to amplify the flanking sequences (5’ and 3’ regions) of the glycoprotein
E (gE) gene of BHV-5 resulting in a gE’ vector after sequencial subcloning
steps, and also amplified the enhanced green flourescent protein (EGFP)
reporter gene, subcloned between 5’ and 3’ gE DNA fragments resulting in a gE-EGFP
vector. The method
could be used to construct plasmids for other recombinant
DNA viruses.
KEYWORDS: PCR, Cloning, Plasmid, EGFP, BHV-5,
Bovine herpesvirus.