Reactivation of an integrated disabled viral vector using a Cre-loxP recombination system in Arabidopsis thaliana.
We developed an inactivated DNA replicon of Turnip Mosaic Virus (TuMV), which was reactivated by a recombination event based on the Cre-loxP system. Viral replication was prevented by the insertion of a translation terminator sequence flanked by two loxP sites at the junction of the P1-HCPro-coding genes. In vitro recombination was tested with purified Cre, which excised the floxed sequence from the TuMV DNA, leaving a single loxP site in the reactivated viral genome, and restored the open reading frame of the replicon. Arabidopsis thaliana plants were made transgenic for the inactivated TuMV replicon. Removal of the translation terminator sequence was achieved by the controlled expression of Cre. Delivery of the Cre recombinase to the transgenic plants was obtained by three methods: agroinfiltration, PVX-based production, or transgenic chemical-inducible expression. In each case, reactivation of TuMV replication was observed.
Tremblay
Arianne
A
Centre de foresterie des Laurentides, Ressources naturelles du Canada, Services canadien des forêts, 1055 rue du PEPS, G1V 4C7, Ste-Foy, QC, Canada.
Beauchemin
Chantal
C
Séguin
Armand
A
Laliberté
Jean-François
JF
D017360:Arabidopsis / Q000235:genetics*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D019426:Integrases / Q000235:genetics; D030821:Plants, Genetically Modified; D017800:Potyvirus
C045073:Cre recombinase; D019426:Integrases