The coat protein of satellite panicum mosaic virus (SPMV) was used to stabilize
viral vector gene inserts in planta. A Potato virus X (PVX) vector carrying the SPMV
capsid protein (CP) gene was successfully stabilized through three serial passages in
Nicotiana benthamiana from the upper non-inoculated leaves following rub inoculation.
The presence of SPMV CP expression from the PVX vector was confirmed by necrotic
lesions that occur only when SPMV CP is present and by western blot and reversetranscription
PCR analyses. In addition, PVX-SPCP was co-inoculated onto N.
benthamiana with a Tomato bushy stunt virus vector carrying a green fluorescent protein
gene, which normally does not yield GFP expression in upper tissue due to loss of the
insert. However, upon co-inoculation with PVX-SPCP, upper non-inoculated leaves
exhibited GFP accumulation based on green fluorescence by UV illumination at 488 nm
and western blot analysis. GFP expression was more abundant in upper non-inoculated
N. benthamiana leaves as well as systemic tissues when the co-inoculation experiments
were performed at 20°C compared to 25°C. These results suggest that SPMV CP is a
viable molecular tool for stabilizing viral vector gene inserts in planta.
| Identifer | oai:union.ndltd.org:tamu.edu/oai:repository.tamu.edu:1969.1/ETD-TAMU-3043 |
| Date | 15 May 2009 |
| Creators | Everett, Anthany Laurence |
| Contributors | Scholthof, Karen-Beth G. |
| Source Sets | Texas A and M University |
| Language | en_US |
| Detected Language | English |
| Type | Book, Thesis, Electronic Thesis, text |
| Format | electronic, application/pdf, born digital |
Page generated in 0.0021 seconds