title: Expression of wild-type and mutant p53 proteins by recombinant vaccinia viruses.
abstract: To facilitate the purification of wild type p53 protein, we established a recombinant p53 vaccinia viral expression system. Using this efficient eukaryotic expression vector, we found that the expressed p53 proteins retained their specific structural characteristics. A comparison between wild type and mutant p53 proteins showed the conservation of the typical subcellular localization and the expression of specific antigenic determinants. Furthermore, wild type p53 exhibited a typical binding with large T antigen, whereas no binding was detected with mutant p53. Both wild type and mutant p53 proteins were highly stable and constituted 5-7% of total protein expressed in the infected cells. These expression recombinant viruses offer a simple, valuable system for the purification of wild type and mutant p53 proteins that are expressed abundantly in eukaryotic cells.
authors.affiliation: Department of Human Microbiology, Sackler School of Medicine, Tel Aviv University, Ramat Aviv, Israel.
authors.forename: D
authors.initials: D
authors.lastname: Ronen
authors.forename: Y
authors.initials: Y
authors.lastname: Teitz
authors.forename: N
authors.initials: N
authors.lastname: Goldfinger
authors.forename: V
authors.initials: V
authors.lastname: Rotter
mesh_terms: D000957:Antigens, Viral, Tumor / Q000378:metabolism; D003001:Cloning, Molecular / Q000379:methods*; D005455:Fluorescent Antibody Technique; D006367:HeLa Cells; D006801:Humans; D009154:Mutation / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D014616:Vaccinia virus / Q000235:genetics*
chemical_list: D000957:Antigens, Viral, Tumor; D011994:Recombinant Proteins; D016159:Tumor Suppressor Protein p53