High level expression in 293 cells of the herpes simplex virus type 2 ribonucleotide reductase subunit 2 using an adenovirus vector.
J Gen Virol
; 71 ( Pt 8): 1785-92, 1990 Aug.
Article
em En
| MEDLINE
| ID: mdl-2167932
ABSTRACT
The herpes simplex viruses (HSV-1 and HSV-2) encode a ribonucleotide reductase consisting of two non-identical subunits (RR1 and RR2) which associate to form the active holoenzyme. To facilitate the purification and subsequent biochemical characterization of this enzyme, we have cloned the small subunit 2 of the HSV-2 ribonucleotide reductase (RR2HSV-2) in a helper-independent adenovirus type 5 vector under the control of the adenovirus type 2 major late promoter. After infection of 293 cells with the recombinant virus, the amount of RR2HSV-2 protein produced was eightfold higher than in HSV-2-infected cells. The specific activities of the RR2HSV-2 recombinant subunit and the RR2HSV-2 protein in HSV-2-infected cells were determined by their mixing with saturating amounts of isolated RR1HSV-1 subunit. By comparison of the relative amount of each RR2HSV-2 subunit with its specific activity, we calculated that the recombinant protein intrinsic activity was similar to that of the protein produced in HSV-2-infected cells. These results demonstrated that the adenovirus expression vector is a good system to produce an active RR2HSV-2 subunit in fairly high amounts.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Ribonucleotídeo Redutases
/
Regulação Viral da Expressão Gênica
/
Simplexvirus
Idioma:
En
Revista:
J Gen Virol
Ano de publicação:
1990
Tipo de documento:
Article
País de afiliação:
Canadá