Figure 6. mLANA mutants that have lost the capacity to repress transcription from the MHV68 TR-driven luciferase reporter construct (MHV68-73TRN mutants) display similar growth kinetics and virus output as the mLANA null mutant virus (73.Stop virus).

Six mLANA mutants were introduced into the MHV68 genome using BAC recombineering. (A) After infection of NIH3T3 fibroblasts with each MHV68-73TRN virus at an MOI of 3 PFU/mL, cell lysates were harvested at one and seven hours post-infection. The lysates were immunoblotted with rabbit mLANA antiserum, mouse MHV68 antiserum, and anti-β-actin antibodies. (B) NIH3T3 fibroblasts were infected at an MOI of 0.001 PFU/cell, and cells and supernatants were harvested at day 7 post-infection and titered on NIH3T12 cells. The resulting data shows that the MHV68-73TRN viruses grow to titers similar to that of 73.Stop, about 10-fold lower than the genetically-repaired 73galkMR virus. **, P<0.01.