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Journal of Virology, June 2006, p. 6182-6187, Vol. 80, No. 12
0022-538X/06/$08.00+0     doi:10.1128/JVI.02447-05
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Efficient In Vitro System of Homologous Recombination in Brome Mosaic Bromovirus{dagger}

Rafal Wierzchoslawski 1,{ddagger} and Jozef J. Bujarski1,2*

Plant Molecular Biology Center and the Department of Biological Sciences, Northern Illinois University, De Kalb, Illinois 60115,1 Institute of Bioorganic Chemistry, Polish Academy of Sciences, Noskowskiego 12/14, 61-704 Poznan, Poland2

Received 21 November 2005/ Accepted 26 March 2006

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.

* Corresponding author. Mailing address: Plant Molecular Biology Center and the Department of Biological Sciences, Northern Illinois University, De Kalb, IL 60115. Phone: (815) 753-0601. Fax: (815) 753-7855. E-mail: jbujarski{at}niu.edu.

{dagger} This is publication number 126 from the Plant Molecular Biology Center at Northern Illinois University.

{ddagger} Present address: HIV Drug Resistance Program, National Cancer Institute at Frederick, National Institutes of Health, Frederick, MD 21702-1201.

Journal of Virology, June 2006, p. 6182-6187, Vol. 80, No. 12
0022-538X/06/$08.00+0     doi:10.1128/JVI.02447-05
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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