Expression of wild-type and mutant p53 proteins by recombinant vaccinia viruses.
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.
Ronen
D
D
Department of Human Microbiology, Sackler School of Medicine, Tel Aviv University, Ramat Aviv, Israel.
Teitz
Y
Y
Goldfinger
N
N
Rotter
V
V
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*
D000957:Antigens, Viral, Tumor; D011994:Recombinant Proteins; D016159:Tumor Suppressor Protein p53