Monitoring viral DNA release with capillary electrophoresis.
Viral DNA injection into host cells is one of the primary mechanisms of viral propagation. Drug development that targets viral propagation requires fast and sensitive methods for monitoring the release of viral DNA in vitro. Here we demonstrate the use of capillary electrophoresis (CE) for monitoring DNA release from virus particles. As a model for this study, we used T5 bacteriophages that infect the bacterium Escherichia coli K-12 by binding to the outer membrane FhuA receptor and then injecting DNA. DNA release from the T5 phages in vitro was induced by either elevated temperature or by interaction with the purified FhuA receptor. After DNA release, the viral samples were stained with the high affinity fluorescent dye YOYO-1, injected into the capillary and subjected to electrophoresis. YOYO-1-stained DNA generated a well-defined peak, allowing reliable detection of viral DNA from as few as 10(5) viral particles. The staining to track T5 phage DNA release exemplifies the great versatility that CE offers in studying viral systems. This CE-based method can be used to study molecular mechanisms of viral infections and to evaluate anti-viral drug candidates.
Krylova
Svetlana M
SM
Department of Chemistry, York University, Toronto, Ontario, Canada. skrylov@yorku.ca
Rozenberg
Dmitry
D
Coulton
James W
JW
Krylov
Sergey N
SN
D004279:DNA, Viral / Q000032:analysis*; D019075:Electrophoresis, Capillary / Q000379:methods; D017201:Virus Shedding
D004279:DNA, Viral