プレキャストGradient Gelを用いた非RI PCR-SSCP法によるp53遺伝子変異の検出

Abstract

We optimized the convenient nonradioactive polymerase chain reaction (PCR)-based single strand conformation polymorphism (SSCP) analysis using commer- cially available precast gradient polyacrylamide gels to detect point mutations in the p53 gene. The optimizatiOn was made by using 2-15% gradient gel whose running conditions were 2×Tris glycine or 1×TBE buffer, and 200 V for 2 hours at 4℃. Also, we screened 47 patients with various hematologic malignancies and detected p53 mutations in two patients with acute leukemia. Both of them showed a resistance to chemotherapy, whereas 20 Other patients with acute leukemia and without detectable mutation did not show such a resistance except for one single case. Mutations in the p53 gene could be closely associated with a resistance to chemotherapy in leukemias.