1995 Fiscal Year Final Research Report Summary
Molecular analysis of congenital C9 deficiency
| Project/Area Number |
06670770
|
|---|
| Research Category |
Grant-in-Aid for General Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
Pediatrics
|
|---|
| Research Institution | The University of Tokyo |
|---|
Principal Investigator |
IGARASHI Takashi The University of Tokyo, Faculty of Medicine, Mejirodai Campus, 医学部・附属病院(分), 講師 (70151256)
|
|---|
| Project Period (FY) |
1994 – 1995
|
| Keywords | Congenital complement component deficiency / Complement 9 (C9) / Molecular analysis |
|---|
| Research Abstract |
1) Northern analysis : Northern analysis of 8 unrelated patients with C9 deficiency was performed resulting with undetected C9 mRNA in two patients.
2) Southern analysis : Southern analysis revealed no major gene rearrangement in all patients with the disease.
3) Ampulification of C9 exons and SSCP analysis :
a) Each exon was amplified by PCR in 2 pateints without C9 mRNA.Each patient revealed to have G to T change in the boundary site of exon 4 suggesting that the splicing of intron 4 was distrubed and synthesis of C9 protein stopped in the intron.
b) SSCP analysis showed that each patient with C9 mRNA had abnormal exon 7.
4) DNA analysis of abnormal exon 7 : DNA sequence of exon 7 revealed a missense mutaion in codon 360. The base change was GGG to GAG suggesting the amino acid change from glycin to glutamic acid in the cell penetrating portion of C9 protein.
5) Oligo-DNA analysis of parents of the 6 patients with G to A change : Oligo-DNAs complementary to normal or wild type were hybidized to DNA of the parents. Both oligo-DNAs were hybridized to the DNA of parents suggesting that the parents were carriers of the same mutation (heterozygote).
|
