Summary
Allelic sequence variation in the apolipoprotein (apo) E gene has been analysed by means of synthetic oligonucleotide probes that detect single base pair substitutions in the codons for amino acid positions 112 and 158, substitutions that are responsible for the common isoforms. Use of the polymerase chain reaction procedure to amplify a sequence of 330 base pairs of the human apo E gene has permitted the development of a robust method for apo E genotyping. This technique has been used to determine the apo E genotype in 95 individuals in whom the genotype for an apo CII TaqI restriction fragment length polymorphism has also been determined. No strong linkage disequilibrium between the two gene loci was detected. This suggests that the metabolic effects of variation, in the apo E and apo CII genes, as detected by the polymorphisms used here, would operate in a statistically independent manner.
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References
Brown MS, Kovanen PT, Goldstein JL (1981) Regulation of plasma cholesterol by lipoprotein receptors. Science 212:628–635
Chakravarti A, Buctow KH, Antonarakis SE, Waber PG, Boehm CD, Kazazian HK (1984) Non-uniform recombination within the human B globin gene cluster. Am J Hum Genet 36:1239–1258
Das HK, McPherson J, Burns GAP, Karathanasis SK, Breslow JL, (1985) Isolation characteristics and mapping to chromosome 19 of the human apolipoprotein E gene. J Biol Chem 260:6240–6247
Davignon J, Gregg RF, Sing CF (1988) Apolipoprotein E polymorphism and atherosclerosis. Arteriosclerosis 8:1–21
Davison PJ, Norton P, Wallis SC, Gill L, Cook M, Williamson R, Humphries SE (1986) There are two gene sequences for human apolipoprotein CI (apo CI) on chromosome 19, one of which is 4kb from the gene for apo E. Biochem Biophys Res Commun 136:876–884
Funke H, Rust S, Assmann G (1986) Detection of apolipoprotein E variants by oligonucleotide “melting” procedure. Clin Chem 32: 1285–1289
Humphries SE, Berg K, Gill L, Cummings AM, Robertson AFH, Stalenhoff A, Williamson R, Borresen AL (1984) The gene for apolipoprotein CII is closely linked to the gene for apolipoprotein E on chromosome 19. Clin Genet 26:389–396
Jackson RL, Baker HN, Gilliam EB, Gotto AM (1977) Primary structure of very low density apolipoprotein CII from human plasma. Proc Natl Acad Sci USA 74:1942–1945
Kessling AM, Horsthemke B, Humphries SE (1985) A study of DNA polymorphisms around the human apolipoprotein A1 gene in hyperlipidaemia and normal individuals. Clin Genet 28:296–306
Klasen EC, Talmud PJ Havekes, L, Wit E de, Van Der Kooij-Meijs E, Smit M, Hansson G, Humphries SE (1987) A common restriction fragment length polymorphism of the human apolipoprotein E gene and its relationship to type III hyperlipidaemia. Hum Genet 75:244–247
Kostner G, Holasek A (1972) Characterisation and quantification of the apolipoproteins from human chyle chylomicrons. Biochemistry 11:1217–1223
La Rosa JC, Levy RI, Herbert P, Lux SE, Fredrickson DS (1970) Specific apolipoprotein activation for lipoprotein lipase. Biochem Biophys Res Commun 41:57–62
Lusis AJ, Heinzmann C, Sparks RS, Scott J, Knott TJ, Geller R, Sparkes MC, Mohandes T (1986) Regional mapping of human chromosome 19: organisation of genes for plasma lipid transport (apo C1 C2 and E and LDL-R) and the genes C3 PEP D and GPI. Proc Natl Acad Sci USA 83:3929–3933
Mahley RW (1988) Apolipoprotein E: cholesterol transport protein with expanding role in cell biology. Science 240:622–630
Maniatis T, Fritsch EF, Sambrook J (eds) (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, pp 122–123
Myklebost OR, Rogne S (1988) A physical map of the apolipoprotein gene cluster on chromosome 19. Hum Genet 78:244–247
Myklebost OR, Williamson R, Mackhom AF, Myklebost S, Rogers J, Woods DE, Humphries SE (1984) The isolation and characterisation of cDNA clones for human apolipoprotein CII. J Biol Chem 259:4401–4404
Olaissen B, Teisberg P, Gedde-Dahl Jr T (1982) The locus of apolipoprotein (apo E) is linked to the complement component 3 (C3) locus on chromosome 19 in man. Hum Genet 62:233–236
Rall SC, Weisgraber KH, Mahley RW (1982) Human apolipoprotein E: the complete amino acid sequence. J Biol Chem 257:4171–4178
Saiki RK, Bugawan TL, Horn GT, Mullis KB, Erlich HA (1986) Analysis of enzymatically amplified β-globin and HLA DQα DNA with allele specific oligonucleotide probes. Nature 324:163–166
Scott J, Knott TJ, Shaw DJ, Brook JD (1985) Localisation of genes encoding apolipoproteins CI, CII and E to p13cen region of human chromosome 19. Hum Genet 71:144–146
Sing CF, Davignon J (1985) Role of the apolipoprotein E polymorphism in determining normal plasma lipid and lipoprotein variation. Am J Hum Genet 37:268–285
Smeets HJM, Poddighe J, Stuyt PMJ, Stalenhoef AFH, Ropers HH, Wieringa B (1988) Identification of apolipoprotein E polymorphism by using synthetic oligonucleotide. J Lipid Res 29:1231–1237
Smit M, Van Der Kooij-Meijs E, Frants RR, Havekes L, Klasen EC (1988) Apolipoprotein gene cluster on chromosome 19. Definitive localisation of the apo C2 gene and the polymorphic Hpal site associated with type III hyperlipoproteinaemia. Hum Genet 78:90–93
Smit M, Van Der Kooij-Meijs E, Van Der Zee A, Van Der Levven G, Stalenhoef A, Havekes L, Frants R (1989) DNA polymorphisms in the apo E-CI-CII gene cluster and the apo B gene in relationship to familial dysbetalipoproteinaemia. Hum Genet (submitted)
Sokal RR, Rholf FJ (1983) Biometry, 2nd edn. Freeman, San Francisco
Thein SL, Wallace RB (1986) The use of synthetic oligonucleotides as specific hybridisation probes in the diagnosis of genetic disorders. In: Davies KE (ed) Human genetic diseases. IRL Press, Oxford Washington, pp 33–50
Thompson EA, Deeb S, Walker D, Motulsky AG (1988) The detection of linkage disequilibrium between closely linked markers. RFLPs at the AI-CIII apolipoprotein genes. Am J Hum Genet 113-124
Utermann G, Hess M, Steinmetz A (1977) Polymorphism of apolipoprotein E and occurrence of dysbetalipoproteinaemia in man. Nature 269:604–607
Wallis SC, Rogne S, Gill L, Markham A, Edge M, Woods D, Williamson R, Humphries SE (1983) The isolation of cDNA clones for human apolipoprotein E and deletion of apo E RNA in hepatic and extrahepatic tissues. EMBO J 12:2369–2373
Weisgraber KH, Newhouse YM, Mahley RW (1988) Apolipoprotein E genotyping using polymerase chain reaction and allele specific oligonucleotide probes. Biochem Biophy, Res Commun 157: 1212–1217
Zannis VI, Breslow JL (1980) Characterisation of the unique human apolipoprotein E variant associated with type III hyperlipoproteinemia. J Biol Chem 225:1759–1762
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Houlston, R.S., Snowden, C., Green, F. et al. Apolipoprotein (apo) E genotypes by polymerase chain reaction and allele-specific oligonucleotide probes: no detectable linkage disequilibrium between apo E and apo CII. Hum Genet 83, 364–368 (1989). https://doi.org/10.1007/BF00291382
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DOI: https://doi.org/10.1007/BF00291382