Abstract
Patients with Duchenne and Becker muscular dystrophy (DMD/BMD) carry mutations in the dystrophin gene. To date, the dystrophin gene is the largest gene ever found in a living organism, measuring 2.4 Mb (1–3). The major muscle transcript consists of 79 exons, spliced together in a 14-kb mature RNA (1,2). The protein coding region spans 11,058 bp, and encodes a 3685 amino acid protein with a mol wt of 427 kDa (2). Transcripts have been identified in many nonmuscular tissues, initiated from eight different promoters, some of which are tissue-specific. Extensive alternative splicing, especially toward the 3′ end of the gene (i.e., exons 70–78), produces a range of slightly shorter transcripts. The gene thus encodes a very complex set of protein isoforms, of which the major constituents have been designated Dp427m (muscle), Dp427c (cortical), Dp427p (Purkinje cells), Dp260 (retina), Dp140 (central nervous system), Dp116 (peripheral nerve), Dp71, and Dp40 (4).
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Monaco, A. P., Neve, R. L., Colletti-Feener, C., Bertelson, C. J., Kurnit, D. M., and Kunkel, L. M. (1986) Isolation of candidate cDNAs for portions of the Duchenne muscular dystrophy gene. Nature 323, 646–650.
Koenig, M., Hoffman, E. P., Bertelson, C. J., Monaco, A. P., Feener, C. A., and Kunkel, L. M. (1987) Complete cloning of the Duchenne muscular dystrophy (DMD) cDNA and preliminary genomic organization of the DMD gene in normal and affected individuals. Cell 50, 509–517.
Den Dunnen, J. T., Grootscholten, P. M., Bakker, E., Blonden, L. A. J., Ginjaar, H. B., Wapenaar, M. C., Van Paassen, H. M. B., et al. (1989) Topography of the DMD gene: FIGE and cDNA analysis of 194 cases reveals 115 deletions and 13 duplications. Am. J. Hum. Genet. 45, 835–847.
Den Dunnen, J. T. and Bakker, E. (1997) The Leiden Muscular Dystrophy data pages. http://www.dmd.nl/.
Chamberlain, J. S., Gibbs, R. A., Ranier, J. E., Nga Nguyen, P. N., and Caskey, C. T. (1988) Deletion screening of the Duchenne muscular dystrophy locus via multiplex DNA amplification. Nucl. Acids Res. 23, 11,141–11,156.
Beggs, A. H., Koenig, M., Boyce, F. M., and Kunkel, L. M. (1990) Detection of 98-percent DMD/BMD gene deletions by polymerase chain reaction. Hum. Genet. 86, 45–48.
Hu, X., Ray, P. N., Murphy, E., Thompson, M. W., and Worton, R. G. (1990) Duplicational mutation at the Duchenne muscular dystrophy locus: its frequency, distribution, origin and phenotype/genotype correlation. Am. J. Hum. Genet. 46, 682–695.
Abbs, S. J., Roberts, R. G., Mathew, C. G., Bentley, D. R., and Bobrow, M. (1990) Accurate assessment of intragenic recombination frequency within the Duchenne muscular dystrophy gene. Genomics 7, 602–606.
Nigro, V., Nigro, G., Esposito, M. G., Comi, L. I., Molinari, A. M., Puca, G. A., and Politano, L. (1994) Novel small mutations along the DMD/BMD gene associated with different phenotypes. Hum. Mol. Genet. 3, 1907–1908.
Roberts, R. G., Bobrow, M., and Bentley, D. R. (1992) Point mutations in the dystrophin gene. Proc. Natl. Acad. Sci. USA 89, 2331–2335.
Roest, P. A. M., Roberts, R. G., Sugino, S., Van Ommen, G. J. B., and Den Dunnen, J. T. (1993) Protein truncation test (PTT) for rapid detection of translation-terminating mutations. Hum. Mol. Genet. 2, 1719–1721.
Gardner, R. J., Bobrow, M., and Roberts, R. G. (1995) The identification of point mutations in Duchenne muscular dystrophy patients using reverse transcript PCR and the protein truncation test. Am. J. Hum. Genet. 57, 311–320.
Carter, M. S., Doskow, J., Morris, P., Li, S., Nhim, R. P., Sandstedt, S., and Wilkinson, M. F. (1995) A regulatory mechanism that detects premature nonsense codons in T-cell receptor transcripts in vivo is reversed by protein synthesis inhibitors in vitro. J. Biol. Chem. 270, 28,995–29,003.
Rowan, A. J. and Bodmer, W. F. (1997) Introduction of a myc reporter tag to improve the quality of mutation detection using the protein truncation test. Hum. Mutat. 9, 172–176.
Roberts, R. G., Bentley, D. R., and Bobrow, M. (1993) Infidelity in the structure of ectopic transcripts: a novel exon in lymphocyte dystrophin transcripts. Hum. Mutat. 2, 293–299.
Whittock, N. V., Roberts, R. G., Mathew, C. G., and Abbs, S. J. (1997) Dystrophin point mutation screening using a multiplexed Protein Truncation Test. Genet. Testing 1, 115–123.
Milasin, J., Muntoni, F., Severini, G. M., Bartoloni, L., Vatta, M., Krajinovic, M., et al. (1996) A point mutation in the 5’ splice site of the dystrophin gene first intron responsible for x linked dilated cardiomyopathy. Hum. Mol. Genet. 5, 73–79.
Roberts, R. G. and Bobrow, M. (1998) Dystrophins in vertebrates and invertebrates. Hum. Mol. Genet. 7, 589–595.
Roest, P. A. M., Roberts, R. G., Van Der Tuijn, A. C., Heikoop, J. C., Van Ommen, G. J. B., and Den Dunnen, J. T. (1993) Protein truncation test (PTT) to rapidly screen the DMD-gene for translation-terminating mutations. Neuromusc. Disord. 3, 391–394.
Roest, P. A. M., Bout, M., Van Der Tuijn, A. C., Ginjaar, H. B., Bakker, E., Hogervorst, F. B. L., Van Ommen, G. J. B., and Den Dunnen, J. T. (1996) Splicing mutations in DMD/BMD detected by RT-PCR/PTT: detection of a 19AA insertion in the cysteine-rich domain of dystrophin compatible with BMD. J. Med. Genet. 33, 935–939.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2001 Humana Press Inc.,Totowa, NJ
About this protocol
Cite this protocol
den Dunnen, J.T. (2001). Point Mutation Detection in the Dystrophin Gene. In: Bushby, K.M., Anderson, L.V. (eds) Muscular Dystrophy. Methods in Molecular Medicine, vol 43. Springer, Totowa, NJ. https://doi.org/10.1385/1-59259-138-8:85
Download citation
DOI: https://doi.org/10.1385/1-59259-138-8:85
Publisher Name: Springer, Totowa, NJ
Print ISBN: 978-0-89603-695-6
Online ISBN: 978-1-59259-138-1
eBook Packages: Springer Protocols