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A novel approach for the rapid mutagenesis and directed evolution of the structural genes of West Nile virus

Wednesday, October 26, 2011 — Poster Session IV

2:00 p.m. – 4:00 p.m.

Natcher Conference Center



* FARE Award Winner


  • T-Y Lin
  • KA Dowd
  • CJ Manhart
  • S Nelson
  • SS Whitehead
  • TC Pierson


Molecular clone technology has proven to be an extremely powerful tool for investigating the lifecycle of flaviviruses, their interactions with the host, and during vaccine development. However, the feasibility of employing existing strategies in large-scale mutagenesis efforts is limited by the technical challenges of manipulating the large and unstable molecular clone plasmids that encode these viruses. In this study, we describe a novel strategy that streamlines the production of infectious West Nile virus (WNV), enabling the introduction of mutations into the viral structural genes at an unprecedented scale. The innovative aspect of this approach is that it does not involve traditional cloning steps that are both time-consuming and require the propagation of potentially unstable constructs in bacteria. This system will enable new genetic approaches to study the structure, function, and immunogenicity of the structural proteins of flaviviruses.

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