The Anti-interferon Activity of Conserved Viral dUTPase ORF54 is Essential for an Effective MHV-68 Infection
Figure 2
Generation of a dUTPase-null MHV-68 ORF54.
A) Immunoblot against the FLAG epitope of immunoprecipitated ORFs or control demonstrates equal amounts of each protein was purified from transfected 293T cells and used in the dUTPase activity assay. B) A PCR reaction utilizing individual dNTPs, with dUTP replacing dTTP, was used to determine the presence of functional dUTP after incubation (between 0 and 25 hours) with purified murine cellular dUTPase, MHV-68 ORF 54, MHV-68 ORF54 H80A/D85N, and a vector control. The presence of a PCR product indicates intact dUTP, and thus a non-functional dUTPase.