FIG. 2.

Design and confirmation of recombinant B virus harboring mutations in thymidine kinase. (A) Overview of TK mutant construction: 1, orientation of the B virus genome; 2, BamHI-SalI fragment containing the complete TK open reading frame; 3, ApaI-BspEI collapse of pBVTK, deleting 754 amino acids from the center of TK; 4, stop codon linker insertion into the BglII site to prematurely stop TK translation. (B) Confirmation of mutants by PCR. Lanes: M, φX174/HaeIII marker; 1, E2490; 2, BVΔTK; 3, BVTKstop PCR product cut with SpeI; 4, BVTKstop (uncut). (C) Antiviral sensitivities of E2490, BVΔTK, and BVTKstop versus acyclovir, ganciclovir, and vidarabine. Error bars indicate standard deviations of three replicates.