The molecular characterization of an A:T to G:C transition in theHbb-b1 gene of the murine homologue of hemoglobin Rainier
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AnN-ethyl-N-nitrosourea (ENU)-induced mutation in theHbb-b1 gene of the mouse hemoglobin-beta complex (Hbb) has been shown to result in a high-oxygen affinity hemoglobin, homologous with hemoglobin Rainier in man (Peters, J.,et al., Genetics 110:709, 1985). Substitution of β145 tyrosine by cysteine had occurred in both human and mouse forms, probably as the result of a point mutation. Provided that sufficient sequence information is available, point mutations can be directly and rapidly analyzed by allele-specific amplification (ASA), as this technique is sensitive enough to detect single nucleotide differences. We report the use of ASA to detect and characterize the mutation in the murine β-globin gene,Hbb-b1d-m1, and find that the codon for β145 tyrosine (TAC) has been replaced by the codon for cysteine (TGC). Therefore, ENU induced an A:T → G:C transition.
Key wordsmouse β-globin complex N-ethyl-N-nitrosourea-induced mutation allele-specific amplification
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- Eckert, K. A., Ingle, C. A., Klinedist, T., and Drinkwater, N. R. (1988). Molecular analysis of mutations induced in human cells by N-ethyl-N-nitrosourea.Mol. Carcinogen. 150.Google Scholar
- Fischel-Ghardsian, N., Hirsh, P. C., and Bohlman, M. C. (1990). Rapid detection of the haemoglobin C mutation by allele specific polymerase chain reaction.Am. J. Hum. Genet. 471023.Google Scholar
- Gilmam, J. G. (1976). Mouse haemoglobin beta chains. Comparative sequence data on adult major and minor beta chains from two species,Mus musculus andMus cervicolor.Biochem. J. 159294.Google Scholar
- Jones, J., and Peters, J. (1991). An ENU induced mutation in the β-globin geneHbb-b1 results in the loss of anRsaI site.Mouse Genome (in press).Google Scholar
- Kwok, S., Kellog, D. E., McKinney, N., Spasic, D., Gola, L., Levinson, C., and Sninsky, J. J. (1990). Effects of primer-template mismatches on the polymerase chain reaction: Human immunodeficiency virus type I model studies.Nucl. Acid Res. 18999.Google Scholar
- Lewis, S. E., Johnson, F. M., Skow, L. C., Popp, D., Barnett, L. B., and Popp, R. A. (1985). A mutation in the mouse beta-globin gene detected in the progeny of a female mouse treated with ethylnitrosourea.Proc. Natl. Acad. Sci. USA 825828.Google Scholar
- Newton, C. R., Graham, A., Heptinstall, L. E., Powell, S. J., Summers, C., Kalsheker, N., Smith, J. C., and Markham, A. F. (1989). Analysis of any point mutation in DNA: The amplification refractory mutation system, ARMS.Nucl. Acid Res. 172503.Google Scholar
- Pastinick, A., Vreeken, C., Nivaard, M. J. M., Searles, L. L., and Vogel, E. W. (1989). Sequence analysuis of N-ethyl-N-nitrosourea induced vermillion mutations inDrosophila melanogaster.Genetics 123485.Google Scholar
- Peters, J., Jones, J., Ball, S. T., and Clegg, J. B. (1990). Analysis of electrophoretically detected mutations induced in mouse germ cells by ethylnitrosourea.Banbury Report 34: Biology of Mammalian Germ Cell Mutagenesis; p. 247.Google Scholar