Efficient in vitro system of homologous recombination in brome mosaic bromovirus.
Recent in vivo studies have revealed that the subgenomic promoter (sgp) in brome mosaic bromovirus (BMV) RNA3 supports frequent homologous recombination events (R. Wierzchoslawski, A. Dzianott, and J. Bujarski, J. Virol. 78:8552-8564, 2004). In this paper, we describe an sgp-driven in vitro system that supports efficient RNA3 crossovers. A 1:1 mixture of two (-)-sense RNA3 templates was copied with either a BMV replicase (RdRp) preparation or recombinant BMV protein 2a. The BMV replicase enzyme supported a lower recombination frequency than 2a, demonstrating a role of other viral and/or host factors. The described in vitro system will allow us to study the mechanism of homologous RNA recombination.
Wierzchoslawski
Rafal
R
Plant Molecular Biology Center, Department of Biological Sciences, Northern Illinois University, De Kalb, IL 60115, USA.
Bujarski
Jozef J
JJ
D017795:Bromovirus / Q000235:genetics*; D002474:Cell-Free System; D003546:Cysteine Endopeptidases / Q000235:genetics; D011401:Promoter Regions, Genetic; D012324:RNA-Dependent RNA Polymerase / Q000235:genetics; D011995:Recombination, Genetic; D013698:Templates, Genetic
D012324:RNA-Dependent RNA Polymerase; D003546:Cysteine Endopeptidases