Studies on Genotyping and Phenotyping of Apolipoprotein E

概要

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We determined the apoE genotypes of 85 healthy donor samples by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Two hundred ninety-two base pair fragments of the exon 4 of the apoE gene were amplified by PCR from genomic DNA extracted from peripheral blood leukocytes, and the PCR products were digested by a restriction endonuclease ; <I>Hha</I> I. The genotype was determined by electrophoresis on a 13 % polyacrylamide gel. These genotypes were compared with the corresponding phenotypes assessed by isoelectric focusing (IEF) and western blotting. The genotypic frequencies were as follows:ε 4/ ε 4, 3.5% ; ε4/ ε 3, 21.2%; ε 4/ ε 2, 2.4%; ε 3/ ε 3, 65.9%; ε 3/ ε 2, 7.1% and ε 2/ ε 2, 0%. The apoE genotypes were completely consistent with the phenotypes obtained by the IEF method. ApoE genotyping by the PCR-RFLP method may not only substitute for phenotyping but may also be useful to detect the variant form of apoE. Therefore, it is a reliable, useful and simple method for analysis of apoE phenotypes and rare polymorphisms.
Japan Society of Clinical Chemistryの論文