Polymerase chain reaction–single-strand conformation polymorphism (PCR-SSCP) analysis of the p53 gene and expression of p53 protein in the leukemic tissues ofBCR/ABLtg/−p53+/− mice.
(A) Schematic model of the region of the mouse p53 gene that showed a polymorphic pattern between the p53 wild-type allele and the p53 null allele. The nucleotide sequences and amino acids of the wild-type allele and the null allele, which showed a polymorphism, are also shown. The box denotes exon 5; black bars, adjacent introns; white triangles, locations of the primers used for amplification (p5-1 and p5-2); and the diamond, the polymorphic site. (B) Results of PCR-SSCP. DNAs extracted from the tumor tissues (T) of 9BCR/ABLtg/−p53+/−leukemic mice (no. 1-9) were subjected to PCR-SSCP analysis. In mice 2, 5, and 8, DNAs extracted from the normal tissues (N) were also analyzed to provide an internal control. DNAs extracted from the normal tissues of a p53+/− mouse and aBCR/ABLtg/− mouse were used as controls for detecting the migration patterns of the parental alleles. Black triangles denote the position of allele A (null); and white triangles, the position of allele B (wild-type). (C) Absence of p53 protein. Proteins extracted from tumor tissues (T) and normal tissues (N) of mice 2, 5, and 8 were subjected to immunoprecipitation/Western blot analysis using anti-p53 antibodies. Normal tissue from a p53+/− mouse was used as a control (denoted by C). The arrow indicates the position of p53.
