Lipoprotein lipase genotypes for a common premature termination codon mutation detected by PCR-med iated site-d i rected mutagenesis and restrict ion digest ion

We have developed a procedure for the determina- tion 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 (Sef14'-Ter) and results in a truncated LPL molecule lacking the C-terminal dipeptide SER-GLY. The mutation can be detected by polymer- ase 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-I11 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 Ser4*'-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.-Stocks, J., J. A. Thorn, and D. J. Galton. Lipo- protein lipase genotypes for a common premature termination codon mutation detected by PCR-mediated site-directed muta- genesis and restriction digestion. J. Lipid Res. 1992. 33: 853- 857.