| Project/Area Number |
09670171
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Pathological medical chemistry
|
|---|
| Research Institution | National Cardiovascular Center Research Institute |
|---|
Principal Investigator |
MORISAKI Takayuki National Cardiovascular Center, Department of Bioscience, Director, バイオサイエンス部, 部長 (30174410)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
MORISAKI Hiroko National Cardiovascular Center, Department of Bioscience, Laborarory Head, バイオサイエンス部, 室長 (40311451)
HIDAKA Kyoko National Cardiovascular Center, Department of Bioscience, Laborarory Head, バイオサイエンス部, 室員 (00216681)
向井 常博 佐賀医科大学, 医学部・生化学, 教授 (40108741)
|
|---|
| Project Period (FY) |
1997 – 1998
|
| Project Status |
Completed (Fiscal Year 1998)
|
| Budget Amount *help |
¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 1998: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1997: ¥1,800,000 (Direct Cost: ¥1,800,000)
|
|---|
| Keywords | AMAP deaminase / purine metabolism / adenine nuleotide / gene targeting / heart / アデニンヌクレオチド / クローニング |
|---|
| Research Abstract |
AMPD gene family is thought to play an important role in purine nucleotide metabolism. To investigate functional roles of AMPD gene family in cellular and animal model, members of mouse Ampd gene family have been isolated and gene targeting experiments have been performed.
First, mouse cDNA for heart-type isoform of AMPD (Ampd3) was isolated, followed by isolation of mouse cDNAs for Ampdl and Ampd3 Then, genomic DNA for corresponding Ampd gene was isolated. Sequence information revealed that mouse Ampd gene indeed functionally corresponds to a member of human AMPD gene family, respectively. Gene targeting is being performed to clarify the relationship between human AMPD1 mutation and metabolic myopathy. In addition, this expriment is expected to help to understand the recent observation that AMPD1 mutation seems to correlate better prognosis of cardiac failure.
Regarding gene targeting, a targeting vector was constructed using corresponding Ampd genomic DNA as well as Neo gene and DT-A gene for positive/negative seletion. Those vectors were introduced into embryonic stem cells and G418 selected colonies were isolated. By PCR-based screening, six homologously recombined clones were establised for Ampd3 and Ampdl. Two clones of them were confirmed to carry normal karyotype. We have already obtained chimeric mice for Ampd3 or Ampdl knock-out. We are currently trying to obtain heterozygous or homozygous mice for Ampd3 or Ampdl gene knock-out. Also, we have identified new AMPD1 mutations in patients with myopaty and are currently doing functional analysis of these mutations. Further investigation of these experiments will give us a new insight of functions of Ampd gene family.
|
