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Generation of individual knockout or tagged mutants of vFLIP or vBcl-2 in the BAC16 genetic system. BAC16-vFLIP-Flag, BAC16-vFLIP-KO, BAC16-vBcl-2-HA, and BAC16-vBcl-2-KO constructs were generated by two-step bacteriophage lambda Red-mediated recombination. (A) BAC16 DNAs isolated from overnight cultures were digested by NheI and subjected to gel electrophoresis. (B) The sequence of each BAC16 mutant clone was confirmed by DNA sequencing. vcyclin, viral cyclin.
