Generating Recombinant Adenoviruses Expressing Cdc25A Variants

To generate recombinant adenoviruses expressing Cdc25A-wild type (wt) or mutants, DNA encoding wt or mutant Cdc25A (S76A or S82A) was, respectively, excised from pCDNA4/TO-Cdc25A-wt, S76A, or S82A [51 (link)] (gifts from Dr. Xianghong Zou, Ohio State University, Columbus, OH, USA) using BamH1 and EcoR1. The recombinant adenoviruses and the control virus expressing green fluorescence protein (GFP) were generated and amplified using ViraPower^™ Adenoviral Gateway^™ Expression Kit (Invitrogen, Carlsbad, CA, USA) following the manufacture's instruction. All adenoviruses were amplified, titrated and used as described previously [52 (link), 53 (link)].