Lipoprotein lipase genotypes for a common premature termination codon mutation detected by PCR-mediated site-directed mutagenesis and restriction digestion.
We have developed a procedure for the determination of a common mutation in exon 9 of the human lipoprotein lipase (LPL) gene. The mutation is due to a C-G transversion which creates a premature termination codon (Ser447-Ter) and results in a truncated LPL molecule lacking the C-terminal dipeptide SER-GLY. The mutation can be detected by polymerase chain reaction (PCR) amplification of exon 9 using a modified 3' amplimer that produces a 140 bp product containing a site for the restriction enzyme Hinf-1 in the presence of the mutation (G allele). The G allele was in strong linkage disequilibrium with a Hind-III restriction fragment length polymorphism (RFLP) allele in intron 8. Genotype determinations for the mutation can be performed by PCR amplification of genomic DNA, digestion with Hinf-1, and analysis of the products by polyacrylamide gel electrophoresis. The allelic frequency of the Ser447-Ter mutation in normal male Caucasian controls was 0.11. The frequency of the mutation was lower in a group of subjects with primary hypertriglyceridemia compared to normolipidemic controls.
Department of Human Genetics and Metabolism, St. Bartholomew's Hospital, London, United Kingdom.
J
J
Stocks
J A
JA
Thorn
D J
DJ
Galton
D000328:Adult; D000368:Aged; D000483:Alleles; D001483:Base Sequence; D003062:Codon; D015247:Deoxyribonuclease HindIII; D005838:Genotype; D005854:Germ Cells / Q000737:chemistry; D006801:Humans; D015228:Hypertriglyceridemia / Q000235:genetics; D008055:Lipids / Q000097:blood; D008071:Lipoprotein Lipase / Q000235:genetics*; D008075:Lipoproteins, HDL / Q000097:blood; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D013728:Terminator Regions, Genetic
D003062:Codon; D008055:Lipids; D008075:Lipoproteins, HDL; D008071:Lipoprotein Lipase; D015247:Deoxyribonuclease HindIII