PZ402, AN IMPROVED SV40-BASED SHUTTLE VECTOR CONTAINING A T-ANTIGEN MUTANT UNABLE TO INTERACT WITH WILD-TYPE P53

Shuttle vectors are useful tools for studying DNA replication and muta genesis. SV40-based shuttle vectors are popular because of their ease of use and quick results. However, one complication with the use of SV 40-based shuttle vectors is the interaction of cellular p53 protein wi th the T-antigen of SV40. Wild-type, but not mutant p53 has been shown to be involved in DNA replication and DNA repair. To address this con cern, we have modified an SV40-based shuttle vector, pZ189, by exchang ing the wt T-antigen for a mutant SV40 T-antigen, which is unable to b ind with p53. This shuttle vector, pZ402, provides us with a tool to s tudy DNA replication and genomic instability in cells with varying gen etic backgrounds without interference from the interaction of T-antige n with p53. (C) 1998 Elsevier Science B.V. All rights reserved.