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Visualization of vaccinia virus infection using the Renilla-luciferase-GFP fusion protein Tatyana Timiryasova*; T Yu; S Shabahang; I Fidor; AA Szalay Center for Molecular Biology & Gene Therapy, Dept. of Biochemistry, Loma Linda, California 92354, USA      *(ifodor@som.llu.edu) The Renilla-luciferase-GFP fusion protein was shown previously to be as a dual reporter in cell culture and in live animals detected by low light image analyses and fluorescence microscopy. Recently we have constructed a recombinant vaccinia virus effectively over-expressing the ruc-gfp cDNA fusion in infected cells. The high yield of protein was confirmed by western immunoblot analysis. A high-resolution reporter gene allows us to monitor replicating viral particles in real time in infected CV-1 and C6 glioma cells by imaging and fluorescence microscopy. Furthermore, the progression of virus infection and dissemination in tumor and different organs of live mice will be assessed using real time imaging system and RG-tagged virus. Finally, the potential use of the RG-tagged vaccinia virus system in tumor gene therapy will be discussed. [Poster: timiryas.tatyan.25632]

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