1996 Fiscal Year Final Research Report Summary
Genetic diagnosis of muscular dystrophy by specific gene amplification
| Project/Area Number |
07557308
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (A)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 試験 |
|---|
| Research Field |
Human genetics
|
|---|
| Research Institution | National Institute of Neuroscience, NCNP |
|---|
Principal Investigator |
ARAHATA Kiichi Department of Neuromuscular Research, National Institute of Neuroscience, NCNP.Director, 神経研究所・疾病研究第一部, 部長 (30053325)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
宋 泯東 国立精神, 神経センター神経研究所・疾病研究第一部, 研究員
HASE Asako Department of Neuromuscular Research, National Institute of Neuroscience, NCNP.D, 神経センター神経研究所・疾病研究第一部, 研究員
SONG Min dong Department of Neuromuscular Research, National Institute of Neuroscience, NCNP.D
|
|---|
| Project Period (FY) |
1995 – 1996
|
| Keywords | Muscular Dystrophy / FSH tyoe / Genetic diagnosis / LA-PCR / Position effect |
|---|
| Research Abstract |
Facioscapulohumeral muscular dystrophy (FSHD) is an autosomal dominant muscular disorder which is characterized by progressive weakness and atrophy of the facial, shoulder-girdle and upper arm muscles, and occasional subsequent pelvic-girdle and lower limb involvement. The gene responsible for FSHD has been localized to chromosome 4q35-qter, although a few chromosome 4-unlinked families are known. Restriction enzyme maps of the polymorphic EcoRI fragment detected by the probes p13E-11 and pFR-1 have revealed that the disease occurs due to a deletion of the integral numbers of the 3.3kb Kpnl tandemly repeated fragments. Deletion mechanism of the LINEs fmily can be studied in the next step.
We have examined 93 Japanese families with possible FSHD.Among these 77 families were confirmed to have 4q35-teromeric rearrangements associated with the disease. Eleven severely affected patients (unrelated) had EcoRI fragment smaller than 11kb. Restriction enzyme maps of the genomic fragments in the
… More
two patients revealed that the 10 kb fragments were identical and contained only one 3.3 kb Kpnl repeat unit. Sequence analysis across the deletion break points showed that the 5' and the 3' elements have the same sequence, and thus suggested the presence of recombination events. We established somatic cell colones from several FSHD patients, which can be provided for detailed chromosome analysis in the teromeric region. To facilitate the genetic diagnosis of FSHD,we also tried quantitative and rapid detection of number of the repeated units within the deleted 4q35 locus by LA-PCR method using pimers franking the repeated units. Successful amplification was accomplished in 84% and each family had a specific LA-PCR amplified product that ranged from 5 to 15 kb in size in which one to four Kpnl repeated units were estimated that were different from the 10q specific repeats. We conclude that the LA-PCR test can be used for accurate and rapid first step screening of deletions of the 4q35-linked FSHD. Less
|
