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

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.
Document type :
Journal articles
Complete list of metadatas

Cited literature [29 references]  Display  Hide  Download

https://hal-anses.archives-ouvertes.fr/hal-00493407
Contributor : Daniel Dory <>
Submitted on : Friday, June 18, 2010 - 4:37:59 PM
Last modification on : Tuesday, October 23, 2018 - 9:54:02 PM
Long-term archiving on : Monday, September 20, 2010 - 5:33:18 PM

File

Author_Manuscript_Faurez_et_al...
Files produced by the author(s)

Identifiers

Collections

Citation

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 Virology Methods, 2010, 165 (1), pp.27-35. ⟨10.1016/j.jviromet.2009.12.013⟩. ⟨hal-00493407⟩

Share

Metrics

Record views

428

Files downloads

430