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CLONING
The study of Duchenne and Becker muscular dystrophy resulted in one of the first
successful attempts at reverse genetics, better described as positional cloning, in humans.
(The other disorder was chronic granulomatous disease, which is also X-linked and
yielded to positional cloning in 1986.) Discovery and subsequent analysis of the gene
mutation that results in the clinical disorder led to the discovery of the encoded protein,
dystrophin. This coinage set a precedent for the naming of proteins discovered by
positional cloning of human disease genes: for example, huntingtin, emerin, and ataxin.
Wood et al. (1987) noted that the gene for Duchenne muscular dystrophy, symbolized
DMD (the same abbreviation as that used for the disorder Duchenne muscular dystrophy)
encodes an mRNA of about 16 kb; thus, if the message encodes a single protein, it would
be about 500 kD in size. The largest proteins in muscle then known were nebulin (550
kD) and titin (over 1000 kD). Both proteins are located at the junction of the A band (the
area of the sarcomere composed of myosin thick filaments) and I band (the area of the
sarcomere containing actin thin filaments attached to the Z line). Wood et al. (1987)
found that the band identified as nebulin was absent or extremely faint in all 30 patients
with DMD whereas it was clear in all controls, and the bands of all other proteins seen in
control muscle were equally prominent in muscle from patients with DMD. Specifically,
titin was equally prominent in control and DMD muscle. The authors suggested that
nebulin may be the defective gene product in DMD; this is now known not to be the case.