Detection of N-ras Mutations in Acute Myeloid Leukemia
N-ras gene activation due to single base substitutions has been found in 20–40% of patients with acute myeloid leukemia (AML) depending on the technique used. Differential hybridization to allele specific oligonucleotide (ASO) probes detects mutations in about 22% of patients (1–6) whereas leukemic DNA assayed by NIH/3T3 transfection ± in vivo transformation in nude mice is associated with a mutation frequency of approximately 40% (7–11). We have developed a rapid screening method, termed allele specific restriction analysis (ASRA), for analysis of mutations at codons 12, 13 and 61 of the N-ras gene (12). A similar approach for the identification of Kirsten-ras gene mutations at codon 12 has also been reported (13,14). ASRA involves PCR amplification of DNA or RNA using a mismatched primer which introduces appropriately positioned base substitutions in N-ras and creates a restriction site provided the adjacent sequence is normal. Resistance of the amplified product to digestion indicates the presence of a mutation in the original template.
KeywordsLymphoma Codon Leukemia Agarose Bromide
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- 8.J.W.G. Janssen, A.C.M. Steenvoorden, J. Lyons, B. Anger, Jv. Bohlke, J.L. Bos, H. Seliger, C.R. Bartram, RAS gene mutations in acute and chronic myelocytic leukemias, chronic myeloproliferative disorders, and myelodysplastic syndromes. Proc. Natl. Acad. Sci. USA 84:9228–9232 (1987).PubMedCrossRefGoogle Scholar
- 13.J. Wei, S.M. Kahn, J.G. Guillem, L. Shih-Hsin, I.B. Weinstein, Rapid detection of ras oncogenes in human tumors: application to colon, esophageal and gastric cancer. Oncogene 4:923–928 (1989).Google Scholar
- 18.A.V. Todd, C.M. Ireland, T.J. Radioff, H. Kronenberg, H.J. Hand, Analysis of N-ras gene mutations in acute myeloid leukemia by allele specific restriction analysis. Am. J. Hematol. (in press) (1991).Google Scholar