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Nucleic Acids Research, 1984, Vol. 12, No. 23 9155-9163
© 1984

MOLECULAR BIOLOGY

Three different mutations in codon 61 of the human N-ras gene detected by synthetic oligonucleotide hybridization

J.L. Bos+, M. Veriaan-de Vries, A.M. Jansen, G.H. Veeneman*, J.H. van Boom* and A.J. van der Eb

Departments of Medical Biochemistry, State University of Leiden, Sytvius Laboratories Wassenaarseweg 72, 2333 AL Leiden, The Netherlands *Organic Chemistry, State University of Leiden, Sytvius Laboratories Wassenaarseweg 72, 2333 AL Leiden, The Netherlands

+To whom correspondence should be sent

Received September 24, 1984. Revised November 2, 1984. Accepted November 2, 1984.

The activation of ras genes in naturally occurring tumors has, thus far, been found to be due to mutations in codon 12 or 61 resulting in single amino acid substitutions. We have used highly labeled synthetic oligonucleo-tides to detect mutations in these codons and to determine the exact position of the mutation. Using this approach we have found three different mutations in codon 61 of the N-ras gene of various human tumor cell lines. In the fibrosarcoma line HT1080 the first nucleotide of the codon is mutated; in the promyelocytic line HL60 the second and in the rhabdomyo-sarcoma line RD301 the third nucleotide. For RD301 this implies that the normal glutamine residue at position 61 is replaced by histidine. In addition to the mutated N-ras gene the three cell lines have a normal N-ras gene which is indicative of the dominant character of the mutations.


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