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Hoffmann, R. A wiki for the life sciences where authorship matters. Nature Genetics (2008)
 

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A functional histidine-tagged replication initiator protein: implications for the study of single-stranded DNA virus replication in planta.

Replication initiation of nanoviruses, plant viruses with a multipartite circular single-stranded DNA genome, is triggered by the master Rep (M-Rep) protein. To enable the study of interactions between M-Rep and viral or host factors involved in replication, we designed oligohistidine-tagged variants of the nanovirus Faba bean necrotic yellows virus (FBNYV) M-Rep protein that allow affinity purification of enzymatically active M-Rep from plant tissue. The tagged M-Rep protein was able to initiate replication of its cognate and other FBNYV DNAs in Nicotiana benthamiana leaf disks and plants. The replicon encoding the tagged M-Rep protein multiplied and moved systemically in FBNYV-infected Vicia faba plants and was transmitted by the aphid vector of the virus. Using the tagged M-Rep protein, we demonstrated the in planta interaction between wild-type M-Rep and its tagged counterpart. Such a tagged and fully functional replication initiator protein will have bearings on the isolation of protein complexes from plants.[1]

References

  1. A functional histidine-tagged replication initiator protein: implications for the study of single-stranded DNA virus replication in planta. Vega-Arreguín, J.C., Timchenko, T., Gronenborn, B., Ramírez, B.C. J. Virol. (2005) [Pubmed]
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