Abstract
While routinely mapping point mutations within the arginase locus of a collection of hyperargininemic patients, we discovered that a base immediately outside a restriction endonuclease recognition site (TaqI) can eliminate cleavage of this site by this enzyme. The genetic lesion lay in a base immediately flanking a TaqI recognition site within exon 8 of the arginase locus and abolished cutting by approximately 80%. We wish to emphasize the necessity of heeding subtle cues frequently encountered while generating restriction enzyme data, because neither Southern blot maps nor endonuclease digestion of polymerase chain reaction amplified products of exon 8 accurately predicted where the point mutation lay. To our knowledge, this is the first instance of inhibition of cleavage by flanking bases occurring on natural (nonsynthetic) D DNA substrates, i.e., within the clinical setting of characterization of a human genetic disorder.
Similar content being viewed by others
Literature cited
The NEB Transcript. (1989). 2(1):8–9.
Frederick, C.A., Grable, J., Melia, M., Samudzi, C., Jen-Jacobson, L., Wang, B.C., Greene, P., Boyer, H.W., and Rosenberg, J.M. (1984).Nature 309327–331.
Kumar, M.R., Hosur, R.V., Roy, K.B., Miles, H.T., and Govil, G. (1985).Biochemistry 247703–7711.
Vinogradova, M.N., Gromova, E.S., Gryaznova, O.I., Isagulyants, M.G., Kuznetsova, S.A., Kosych, V.G., and Shabarova, Z.A. (1987).Bioorg. Khim. 131194–1204.
Grody, W.W., Dodson, A.E., Klein, D., Kern, R. Wissman, P.B., Bassand, P., and Cederbaum, S.D. (1989) (Abstract)Am. J. Hum. Genet. 45:A191.
Brusilow, S.W., and Horwich, A.L. (1989). Urea cycle enzymes. InThe Metabolic Basis of Inherited Disease, 6th ed., (eds.) Scriver, C.R., Beaudet, A.L., Sly, W.S., Valle, D. McGraw-Hill, New York, pp. 629–663.
Cederbaum, S.D., Shaw, K.N.F., Spector, E.B., Verity, M.A., Snodgrass, P.J., and Sugarman, G.I. (1979).Pediatr. Res. 1313827–833.
Bernar, J., Hanson, R.A., Kern, R., Phoenix, B., Shaw, K.N.F., and Cederbaum, S.D. (1986).J. Pediatr. 108432–435.
Hermann, B.G., and Frischauf, A.M. (1987).Methods Enzymol. 152180–183.
Kunkel, L.M., Smith, K.D., Boyer, S.H., Borgaonkar, D.S., Wachtel, S.S., Miller, O.N., Breg, W.R., Jones, H.W. and Rary, J.M. (1977).Proc. Natl. Acad. Sci. U.S.A. 741245–1249.
Dizikes, G.J., Grody, W.W., Kern, R.M., and Cederbaum, S.D. (1986).Biochem. Biophys. Res. Commun. 14153–59.
Southern, E.M. (1975).J. Mol. Biol. 98503–517.
Gatti, R.A., Concannon, P., and Salser, W. (1984).BioTechniques 2148–155.
Saiki, R.K., Gelfand, D.H., Stoffel, B., Scharf, S.J., Higuchi, R., Horn, G.T., Mullis, K.B., Erlich, H. A. (1988).Science 239487–491.
Haraguchi, Y., Takiguchi, M., Amaya, Y., Kawamoto, S., Matsuda, I., and Mori, M. (1987).Proc. Natl. Acad. Sci. U.S.A. 84412–415.
Brow, M.N.D. (1990). Sequencing with Taq DNA polymerase. InPCR Protocols: A Guide to Methods and Applications, Innis, M.A., Gelfand, D.H., Sninsky, J.J., and White, T.J. (eds.) (Academic Press, San Diego), pp. 189–196.
Watson, J.D., Hopkins, N.H., Roberts, J.W., Steitz, J.A., and Weiner, A.M. (1987).Molecular Biology of the Gene, 4th ed. (The Benjamin Cummings Publishing Co., Menlo Park, California), pp. 248–254, 262–270.
Gingeras, T.R., and Brooks, J.E. (1983).Proc. Natl. Acad. Sci. U.S.A. 80402–406.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Klein, D., Dodson, A.E., Tabor, D.E. et al. Effect of an adjacent base on detection of a point mutation by restriction enzyme digestion. Somat Cell Mol Genet 17, 369–375 (1991). https://doi.org/10.1007/BF01233062
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF01233062