Poster: Education

Abs # 32: Overexpression and silencing of genes in transfected Nicotiana benthamiana using a plant RNA viral vector

Presenter:	Kaneko, Osamu F., okaneko@fas.harvard
Authors	Kaneko, Osamu F. (A)   Kumagai, Monto H. (B)
Affiliations:	(A): Harvard University (B): University of Hawaii, Department of Molecular Biosciences & Bioengineering

A summer research opportunity for undergraduate students was made available by support from the National Science Foundation and the Marine Bioproducts Engineering Center (MarBEC). The 10-week MSURF (Marine Science Undergraduate Research Fellowship) program was coordinated through both the University of Hawaii at Manoa and the University of California, Berkeley. The goals of the internship were to provide aspiring researchers with valuable laboratory experience, specifically through work with plant viruses. Using a modified variant of the Tobacco Mosaic Virus (TMV), genes from the marine alga, Dunaliella salina were transported into the tobacco related plant, Nicotiana benthamiana in an effort to achieve post-transcriptional gene silencing. Foreign genes from the marine coral, Discosoma and from the marine alga were also over-expressed within N. benthamiana plants. The gene encoding the fluorescent protein DsRed from Discosoma and a RNA helicase from D. salina, were placed under the transcriptional control of a tobamovirus subgenomic promoter. 1-2 weeks following inoculation with the hybrid viral vectors, N. benthamiana plants were examined for over-expression and silencing of the targeted genes. We have developed methods to determine the global change in gene expression caused by viral derived expression or silencing of genes in transfected plants. This technique can also be a valuable tool for the isolation and characterization of unknown genes.