Table 4.
Determination of human mEH allele classification.
| (a) Allele-specific hybridization conditions | ||
|---|---|---|
| Amino acid position | Hybridization (T°C) | Wash (T°C) |
| Exon 3 | ||
| Tyr/His 113 | 52 | 52 |
| Exon 4 | ||
| His/Arg 139 | ND | ND |
| His/Asn 148 | 60 67.5 | |
| Exon 8 | ||
| Lys/Ser 348 | 54 | 54 |
| Exon 9 | ||
| Thr/Ile 396 | 57 | 57 |
| Leu/Phe 406 | 54 | 54 |
| Leu/Val 420 | 54 | 54 |
|
| ||
| (b) Allele-specific restriction enzymes | ||
| Allelic amino acid | Restriction enzyme sensitivity | |
| Exon 4 | ||
| Arg 139 | RsaI | |
| His 148 | ApaLI | |
| Asn 148 | HphI | |
| Exon 8 | ||
| Ser 348 | HindIII | |
Amino acid variants were genotyped in individuals by hybridization to allele-specific oligonucleotides under defined conditions (a), or, in some cases, by PCR-RFLP analysis of amplified exons (b). Optimal hybridization and wash conditions for allelic variants at residue 139 in exon 4 were not determined (ND)