GM08369
LCL from B-Lymphocyte
Description:
HEMOPHILIA B; HEMB
5,10-@METHYLENETETRAHYDROFOLATE REDUCTASE; MTHFR
Repository
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NIGMS Human Genetic Cell Repository
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Subcollection |
Chromosome Abnormalities |
Class |
X Chromosome Markers |
Biopsy Source
|
Peripheral vein
|
Cell Type
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B-Lymphocyte
|
Tissue Type
|
Blood
|
Transformant
|
Epstein-Barr Virus
|
Sample Source
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LCL from B-Lymphocyte
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Race
|
White
|
Relation to Proband
|
proband
|
Confirmation
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Clinical summary/Case history
|
Species
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Homo sapiens
|
Common Name
|
Human
|
Remarks
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IDENTIFICATION OF SPECIES OF ORIGIN |
Species of Origin Confirmed by Nucleoside Phosphorylase, Glucose-6-Phosphate Dehydrogenase, and Lactate Dehydrogenase Isoenzyme Electrophoresis |
|
Gene |
MTHFR |
Chromosomal Location |
1p36.3 |
Allelic Variant 1 |
607093.0003; MTHFR THERMOLABILE POLYMORPHISM |
Identified Mutation |
677C>T; Frosst et al. [Nature Genet. 10: 111-113 (1995)] identified a C-to-T substitution at nucleotide 677 that converted an alanine to a valine residue. The alteration created a HinfI site that was used to screen 114 unselected French-Canadian chromosomes; the allele frequency of the substitution was 0.38. The mutation in the heterozygous or homozygous state correlated with reduced enzyme activity and increased thermolability in lymphocyte extracts; in vitro expression of the mutagenized cDNA containing the mutation confirmed its effect on thermolability of MTHFR. Individuals homozygous for the mutation had significantly elevated plasma homocysteine levels. Thus, the 677C-T mutation may represent an important genetic risk factor in vascular disease. |
Remarks |
Factor IX deficiency diagnosed at age 15 mos; Factor IX level <1%; hemarthroses involving multiple joints & a massive intramuscular hematoma; prolonged PPT; mother & 2 sisters have normal Factor IX levels; donor subject is heterozygous for a C>T mutation at nucleotide 677 in exon 4 of the methylenetetrahydrofolate reductase (MTHFR) gene [677C>T] that results in a substitution of a valine for an alanine at codon 222 [Ala222Val (A222V)]; donor subject is homozygous normal for the R506Q mutation in the F5 gene and for the 20210G>A variant in the prothrombin gene (F2) |
Split Ratio |
1:5 |
Temperature |
37 C |
Percent CO2 |
5% |
Medium |
Roswell Park Memorial Institute Medium 1640 with 2mM L-glutamine or equivalent |
Serum |
15% fetal bovine serum Not Inactivated |
Substrate |
None specified |
Subcultivation Method |
dilution - add fresh medium |
Supplement |
- |
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