[Specific amplification by PCR for N-ras mutation in thyroid follicular cartinoma].
Mutation-specific PCR amplification, a special version of allele-specific amplification, is a method for detection of known mutation. In this method, one of the paired primers has a base at the 3' terminal that is complementary to a mutated base. With this primer, only alleles which have the same mutation can be amplified. We analyzed point mutations of N-ras protooncogene codon 61 in thyroid neoplasms using this mutation-specific PCR amplification method. This method is easy to perform, and many samples can be treated at one time. We detected 2 point mutations of N-ras codon 61 out of 7 follicular carcinomas. One was a cytosine to adenine substitution mutation at the first base, and the other was an adenine to guanine substitution mutation in the second base. These mutations were not detected in 7 follicular adenoma and 1 papillary carcinoma. The results were confirmed by both dot blot hybridization and direct sequencing. Mutation-specific amplification is a powerful method for detection of known mutations of protooncogenes or tumor suppressor genes in investigation of the cancer biology.
Taniyama
M
M
Third Department of Internal Medicine, Showa University.
Kaihara
M
M
D018263:Adenocarcinoma, Follicular / Q000235:genetics*; D001483:Base Sequence; D003062:Codon; D011905:Genes, ras / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D013964:Thyroid Neoplasms / Q000235:genetics*
D003062:Codon