Replication efficiency of rolling-circle replicon-based plasmids derived from porcine circovirus 2 in eukaryotic cells. - Archive ouverte HAL Access content directly
Journal Articles Journal of Virological Methods Year : 2010

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

(1) , (1) , (1) , (1) , (1)
1

Abstract

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.
Fichier principal
Vignette du fichier
Author_Manuscript_Faurez_et_al_J_Virol_Methods_2010.pdf (307.54 Ko) Télécharger le fichier
Origin : Files produced by the author(s)
Loading...

Dates and versions

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

Identifiers

Cite

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⟩

Collections

ANSES
196 View
528 Download

Altmetric

Share

Gmail Facebook Twitter LinkedIn More