| Project/Area Number |
09670858
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Pediatrics
|
|---|
| Research Institution | Gifu University |
|---|
Principal Investigator |
ORII Tadao Chubu Gakuin University Faculty of Human Welfare, Professor, 人間福祉学部, 教授 (20045339)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
SUKEGAWA Kazuko Gifu University School Medicine, Research Associate, 医学部, 助手 (60115409)
|
|---|
| Project Period (FY) |
1997 – 1999
|
| Project Status |
Completed (Fiscal Year 1999)
|
| Budget Amount *help |
¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 1999: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1998: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1997: ¥1,200,000 (Direct Cost: ¥1,200,000)
|
|---|
| Keywords | mucopolysaccharidosis / GAINS gene cloning / mouse model / mutation analysis / 遺伝性ムコ多糖症 / モルキオ病 |
|---|
| Research Abstract |
In order to facilitate in vivo studies using model animals for MPS IV A, we isolated and performed molecular characterization of the mouse homolog of human GALNS. The 2.3-kbcDNA contains a 1560-bp open reading frame encoding 520 amino acid residues. The coding region has 84%similarity to the human GALNS cDNA at amino acid level. The mouse Galns gene was mapped by interspecific backcross analysis to the distal region of chromosome 8 where it co-segregates with Aprt. Northern blot analysis showed a wide expression of a single-copy gene, being higher especially in liver and kidney. The Galns gene was about 50-kb long and organized into 14 exons and 13 introns. All intron-exon splice junctions conformed to the GA/AC consensus sequence except exon 8/intron 8 junction. Primer extension shows multiple transcription initiation sites between -44 and -75 although major transcription initiation site was observed at -90bp from the ATG codon. The 5'-flanking region lackes canonical TATA and CAAT box sequences, but is G+C rich with 10 GC boxes (potential Sp1 binding sites), characteristic of a housekeeping gene promoter.
We have constructed a plasmid containing the Galns gene fragment in the Neo+TK vector. The targeting vector introduced into the ES cells. Targeted clones were isolated. Targeted ES cells lines were injected into the blastcytes of embryos. At now, highly chimeric male mice are going to obtain.
|
