Publication Type:Journal Article
Source:J Virol, Volume 81, Number 17, p.9339-45 (2007)
ISBN:0022-538X (Print)<br/>0022-538X (Linking)
Keywords:Animals, Aphids/ virology, Baculoviridae/ genetics, Cell Line, Cell Nucleus/virology, DNA, Complementary, Genetic Vectors, Microscopy, Electron, Transmission, Recombination, Genetic, RNA Viruses/ genetics, RNA, Viral/ genetics, Spodoptera/cytology/virology, Virion/ultrastructure
Detailed investigation of virus replication is facilitated by the construction of a full-length infectious clone of the viral genome. To date, this has not been achieved for members of the family Dicistroviridae. Here we demonstrate the construction of a baculovirus that expresses a dicistrovirus that is infectious in its natural host. We inserted a full-length cDNA clone of the genomic RNA of the dicistrovirus Rhopalosiphum padi virus (RhPV) into a baculovirus expression vector. Virus particles containing RhPV RNA accumulated in the nuclei of baculovirus-infected Sf21 cells expressing the recombinant RhPV clone. These virus particles were infectious in R. padi, a ubiquitous aphid vector of major cereal viruses. The recombinant virus was transmitted efficiently between aphids, despite the presence of 119 and 210 vector-derived bases that were stably maintained at the 5' and 3' ends, respectively, of the RhPV genome. The maintenance of such a nonviral sequence was surprising considering that most RNA viruses tolerate few nonviral bases beyond their natural termini. The use of a baculovirus to express a small RNA virus opens avenues for investigating replication of dicistroviruses and may allow large-scale production of these viruses for use as biopesticides.
Pal, Narinder<br/>Boyapalle, Sandhya<br/>Beckett, Randy<br/>Miller, W Allen<br/>Bonning, Bryony C<br/>Research Support, Non-U.S. Gov't<br/>United States<br/>J Virol. 2007 Sep;81(17):9339-45. Epub 2007 Jun 27.