Replication efficiency of rolling-circle replicon-based plasmids derived from porcine circovirus 2 in eukaryotic cells. - Archive ouverte HAL Accéder directement au contenu
Article Dans Une Revue Journal of Virological Methods Année : 2010

Replication efficiency of rolling-circle replicon-based plasmids derived from porcine circovirus 2 in eukaryotic cells.

Résumé

In this study, a method was developed to measure replication rates of rolling-circle replicon-based plasmids in eukaryotic cells. This method is based on the discriminative quantitation of MboI-resistant, non-replicated input plasmids and DpnI-resistant, replicated plasmids. To do so, porcine circovirus type 2 (PCV2) replicon-based plasmids were constructed. These plasmids contained the PCV2 origin of replication, the PCV2 Rep promoter and the PCV2 Rep gene. The results show that the replication rate depends on the length of the PCV2 replicon-based plasmid and not on the respective position of the Rep promoter and the promoter of the gene of interest that encodes the enhanced green fluorescent protein (eGFP). In all cases, it was necessary to add the Rep gene encoded by a plasmid and cotransfected as a replication booster. This method can evaluate the replication potential of replicon-based plasmids quickly and is thereby a promising tool for the development of plasmids for vaccine purposes.

Domaines

Virologie
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Dates et versions

hal-00493407 , version 1 (18-06-2010)

Identifiants

Citer

Florence Faurez, Daniel Dory, Aurélie Henry, Stéphanie Bougeard, André Jestin. Replication efficiency of rolling-circle replicon-based plasmids derived from porcine circovirus 2 in eukaryotic cells.. Journal of Virological Methods, 2010, 165 (1), pp.27-35. ⟨10.1016/j.jviromet.2009.12.013⟩. ⟨hal-00493407⟩

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