Journal: Journal of Virology
Article Title: Dissociation of Rabies Virus Matrix Protein Functions in Regulation of Viral RNA Synthesis and Virus Assembly
Figure Lengend Snippet: RNA synthesis and growth of SAD L16, SAD M(R58G), and SAD M(R58E) viruses. (A) BSR cells were infected with the viruses at an MOI of 2, and after 1 and 2 days of infection, the RNA was isolated from the cells and analyzed in Northern hybridization with N or L probes. The amount of cellular 28S rRNA at 1 day postinfection (dpi) was detected in a second hybridization step with 28S cDNA probe. n.i., not infected. (B) Transcription rates of SAD L16, SAD M(R58G), and SAD M(R58E) viruses. The transcription rates were calculated for each virus after quantitation of the N (1 and 2 dpi) and L (1 dpi) hybridization signals by phosphorimaging and normalization of the transcript levels with the genome RNA levels. The transcription rates are given as percentages (SAD L16 transcription rate, 100%). (C) Levels of virus RNAs in infected cells. The amounts of virus-specific RNAs in infected cells, detected with the N probe, were normalized to that of 28S rRNA. In SAD M(R58G)-infected cells, the level of virus-expressed RNAs increased to 239% (SAD L16 level, 100%), whereas the virus RNA levels in SAD M(R58E)-infected cells decreased to 27%. (D) One-step growth curves. The infectious virus titers were determined in supernatants of identical infection experiments at days 1, 2, and 3 postinfection.
Article Snippet: To detect cellular RNA, a 28S rRNA probe (Ambion) was used.
Techniques: Infection, Isolation, Northern Blot, Hybridization, Quantitation Assay