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OJVRTM
Online Journal of Veterinary Research©
Volume 9 (1) :
57-65, 2005. Redacted 2018.
PCR-based
plasmid vector for recombinant bovine herpesvirus
De
Souza VPa,c, Dellagostin OAb , Roehe PMa,c.
a Centre of Veterinary Research Desidério
Finamor (CPVDF/FEPAGRO), Estrada do Conde, 6000,
Eldorado do Sul, RS, Brazil, 90001-970. b
Centre of Biotechnology, Federal University of Pelotas (UFPel),
PO Box 354, CEP 96001, Pelotas, RS,
Brazil. c Department of
Microbiology, Virology Laboratory, Federal University of Rio Grande do Sul (UFRGS), Porto Alegre, RS, Brazil.
Corresponding author: Vanessa
Felipe de Souza. E-mail: souzavf@yahoo.com.br
Souza VF, Dellagostin
OA, Roehe PM,. PCR-based
plasmid vector for recombinant bovine herpesvirus, Onl
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 the
traditional methods based in viral DNA restriction endonucleases digestion and
cloning. 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, amplified by enhanced green flourescent protein (EGFP) reporter gene, subcloned between 5’ and 3’ gE
DNA fragments resulting in a gE-EGFP
vector. This vector can be used for the production of recombinant BHV-5 from
deleted viral samples. The method could be tested for construction of plasmids
for production of several recombinant DNA viruses.
KEYWORDS:
PCR, Cloning, Plasmid, EGFP, BHV-5, Bovine herpesvirus.