©1994-2009 All Rights Reserved. Online Journal of Veterinary Research. You may not store these pages in any form except for your own personal use. All other usage or distribution is illegal under international copyright treaties. Permission to use any of these pages in any other way besides the before mentioned must be gained in writing from the publisher. This article is exclusively copyrighted in its entirety to OJVR publications. This article may be copied once but may not be, reproduced or re-transmitted without the express permission of the editors.
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.
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.