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. 1986 Feb;83(3):644–648. doi: 10.1073/pnas.83.3.644

Mapping of the human APOB gene to chromosome 2p and demonstration of a two-allele restriction fragment length polymorphism.

L S Huang, D A Miller, G A Bruns, J L Breslow
PMCID: PMC322920  PMID: 3003743

Abstract

ApoB is a large glycoprotein with an apparent molecular mass of 550 kDa on NaDodSO4/PAGE. It is a major constituent of most lipoproteins and plays an important role in their metabolism. Recently, apoB cDNA clones have been isolated from an expression library made with mRNA from a human hepatoma cell line. These clones, which were all 1.5-1.6 kilobases (kb) long and corresponded to the 3' end of apoB mRNA, were used to demonstrate that hepatic apoB mRNA is approximately 22 kb long. In the current report, a probe derived from one of these cDNA clones, pB8, was used for in situ hybridization experiments to map the human gene for apoB, APOB, to the distal half of the short arm of chromosome 2. This probe was also used to analyze somatic cell hybrids and, in agreement with the in situ hybridization studies, concordancy was demonstrated with chromosome 2. In addition, two hybrids with chromosome 2 translocations that contain only the short arm reacted with the pB8 probe. A third hybrid with a complex rearrangement of chromosome 2, which deleted an interstitial region and the tip of the short arm of chromosome 2, did not react. These data indicate that APOB maps to either 2p21-p23 or 2p24-pter. In further studies, DNA from normal individuals, digested with the restriction endonuclease EcoRI and subjected to Southern blot analysis with the pB8 probe, revealed a two-allele restriction fragment length polymorphism (RFLP). The major allele was 11 kb, and the minor allele was 13 kb. The minor allele was present with a frequency of 20-25%. The inheritance of the two alleles was studied in an informative family, and they segregated in a typical autosomal Mendelian fashion. The mapping studies provide the means for understanding the relationship of the APOB locus to others in the human genome, whereas the demonstration of an APOB RFLP increases our ability to assess the role of this locus in determining plasma lipoprotein levels.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Breslow J. L. Human apolipoprotein molecular biology and genetic variation. Annu Rev Biochem. 1985;54:699–727. doi: 10.1146/annurev.bi.54.070185.003411. [DOI] [PubMed] [Google Scholar]
  2. Brook J. D., Shaw D. J., Meredith L., Bruns G. A., Harper P. S. Localisation of genetic markers and orientation of the linkage group on chromosome 19. Hum Genet. 1984;68(4):282–285. doi: 10.1007/BF00292584. [DOI] [PubMed] [Google Scholar]
  3. Bruns G. A., Karathanasis S. K., Breslow J. L. Human apolipoprotein A-I--C-III gene complex is located on chromosome 11. Arteriosclerosis. 1984 Mar-Apr;4(2):97–102. doi: 10.1161/01.atv.4.2.97. [DOI] [PubMed] [Google Scholar]
  4. Bruns G. A., Leary A. C., Eisenman R. E., Bazinet C. W., Regina V. M., Gerald P. S. Expression of ACONS and GALT in man-rodent somatic cell hybrids. Cytogenet Cell Genet. 1978;22(1-6):172–176. doi: 10.1159/000130929. [DOI] [PubMed] [Google Scholar]
  5. Bruns G. A., Mintz B. J., Leary A. C., Regina V. M., Gerald P. S. Human lysosomal genes: arylsulfatase A and beta-galactosidase. Biochem Genet. 1979 Dec;17(11-12):1031–1059. doi: 10.1007/BF00504344. [DOI] [PubMed] [Google Scholar]
  6. Bruns G., Kavathas P., Shiloh Y., Sakai K., Schwaber J., Latt S. A., Herzenberg L. A. The human T cell antigen Leu-2 (T8) is encoded on chromosome 2. Hum Genet. 1985;70(4):311–314. doi: 10.1007/BF00295366. [DOI] [PubMed] [Google Scholar]
  7. Cheung P., Kao F. T., Law M. L., Jones C., Puck T. T., Chan L. Localization of the structural gene for human apolipoprotein A-I on the long arm of human chromosome 11. Proc Natl Acad Sci U S A. 1984 Jan;81(2):508–511. doi: 10.1073/pnas.81.2.508. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Das H. K., McPherson J., Bruns G. A., Karathanasis S. K., Breslow J. L. Isolation, characterization, and mapping to chromosome 19 of the human apolipoprotein E gene. J Biol Chem. 1985 May 25;260(10):6240–6247. [PubMed] [Google Scholar]
  9. Deeb S. S., Motulsky A. G., Albers J. J. A partial cDNA clone for human apolipoprotein B. Proc Natl Acad Sci U S A. 1985 Aug;82(15):4983–4986. doi: 10.1073/pnas.82.15.4983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fojo S. S., Law S. W., Brewer H. B., Jr, Sakaguchi A. Y., Naylor S. L. The localization of the gene for apolipoprotein C-II to chromosome 19. Biochem Biophys Res Commun. 1984 Jul 31;122(2):687–693. doi: 10.1016/s0006-291x(84)80088-1. [DOI] [PubMed] [Google Scholar]
  11. Harper M. E., Saunders G. F. Localization of single copy DNA sequences of G-banded human chromosomes by in situ hybridization. Chromosoma. 1981;83(3):431–439. doi: 10.1007/BF00327364. [DOI] [PubMed] [Google Scholar]
  12. Huang L. S., Bock S. C., Feinstein S. I., Breslow J. L. Human apolipoprotein B cDNA clone isolation and demonstration that liver apolipoprotein B mRNA is 22 kilobases in length. Proc Natl Acad Sci U S A. 1985 Oct;82(20):6825–6829. doi: 10.1073/pnas.82.20.6825. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Humphries S. E., Berg K., Gill L., Cumming A. M., Robertson F. W., Stalenhoef A. F., Williamson R., Børresen A. L. The gene for apolipoprotein C-II is closely linked to the gene for apolipo-protein E on chromosome 19. Clin Genet. 1984 Nov;26(5):389–396. [PubMed] [Google Scholar]
  14. Jackson C. L., Bruns G. A., Breslow J. L. Isolation and sequence of a human apolipoprotein CII cDNA clone and its use to isolate and map to human chromosome 19 the gene for apolipoprotein CII. Proc Natl Acad Sci U S A. 1984 May;81(10):2945–2949. doi: 10.1073/pnas.81.10.2945. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kanda N., Schreck R., Alt F., Bruns G., Baltimore D., Latt S. Isolation of amplified DNA sequences from IMR-32 human neuroblastoma cells: facilitation by fluorescence-activated flow sorting of metaphase chromosomes. Proc Natl Acad Sci U S A. 1983 Jul;80(13):4069–4073. doi: 10.1073/pnas.80.13.4069. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kane J. P. Apolipoprotein B: structural and metabolic heterogeneity. Annu Rev Physiol. 1983;45:637–650. doi: 10.1146/annurev.ph.45.030183.003225. [DOI] [PubMed] [Google Scholar]
  17. Kannel W. B., Castelli W. P., Gordon T. Cholesterol in the prediction of atherosclerotic disease. New perspectives based on the Framingham study. Ann Intern Med. 1979 Jan;90(1):85–91. doi: 10.7326/0003-4819-90-1-85. [DOI] [PubMed] [Google Scholar]
  18. Karathanasis S. K., McPherson J., Zannis V. I., Breslow J. L. Linkage of human apolipoproteins A-I and C-III genes. 1983 Jul 28-Aug 3Nature. 304(5924):371–373. doi: 10.1038/304371a0. [DOI] [PubMed] [Google Scholar]
  19. Karathanasis S. K., Norum R. A., Zannis V. I., Breslow J. L. An inherited polymorphism in the human apolipoprotein A-I gene locus related to the development of atherosclerosis. Nature. 1983 Feb 24;301(5902):718–720. doi: 10.1038/301718a0. [DOI] [PubMed] [Google Scholar]
  20. Karathanasis S. K., Zannis V. I., Breslow J. L. Isolation and characterization of cDNA clones corresponding to two different human apoC-III alleles. J Lipid Res. 1985 Apr;26(4):451–456. [PubMed] [Google Scholar]
  21. Kessling A. M., Horsthemke B., Humphries S. E. A study of DNA polymorphisms around the human apolipoprotein AI gene in hyperlipidaemic and normal individuals. Clin Genet. 1985 Oct;28(4):296–306. doi: 10.1111/j.1399-0004.1985.tb00403.x. [DOI] [PubMed] [Google Scholar]
  22. Latt S. A., Willard H. F., Gerald P. S. BrdU-33258 Hoechst analysis of DNA replication in human lymphocytes with supernumerary or structurally abnormal X chromosomes. Chromosoma. 1976 Aug 17;57(2):135–153. doi: 10.1007/BF00292912. [DOI] [PubMed] [Google Scholar]
  23. Law S. W., Gray G., Brewer H. B., Jr, Sakaguchi A. Y., Naylor S. L. Human apolipoprotein A-I and C-III genes reside in the p11----q13 region of chromosome 11. Biochem Biophys Res Commun. 1984 Feb 14;118(3):934–942. doi: 10.1016/0006-291x(84)91485-2. [DOI] [PubMed] [Google Scholar]
  24. Lee D. M., Valente A. J., Kuo W. H., Maeda H. Properties of apolipoprotein B in urea and in aqueous buffers. The use of glutathione and nitrogen in its solubilization. Biochim Biophys Acta. 1981 Oct 23;666(1):133–146. doi: 10.1016/0005-2760(81)90099-0. [DOI] [PubMed] [Google Scholar]
  25. Lusis A. J., West R., Mehrabian M., Reuben M. A., LeBoeuf R. C., Kaptein J. S., Johnson D. F., Schumaker V. N., Yuhasz M. P., Schotz M. C. Cloning and expression of apolipoprotein B, the major protein of low and very low density lipoproteins. Proc Natl Acad Sci U S A. 1985 Jul;82(14):4597–4601. doi: 10.1073/pnas.82.14.4597. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Malloy M. J., Kane J. P., Hardman D. A., Hamilton R. L., Dalal K. B. Normotriglyceridemic abetalipoproteinemia. absence of the B-100 apolipoprotein. J Clin Invest. 1981 May;67(5):1441–1450. doi: 10.1172/JCI110173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Mantzouranis E. C., Dowton S. B., Whitehead A. S., Edge M. D., Bruns G. A., Colten H. R. Human serum amyloid P component. cDNA isolation, complete sequence of pre-serum amyloid P component, and localization of the gene to chromosome 1. J Biol Chem. 1985 Jun 25;260(12):7752–7756. [PubMed] [Google Scholar]
  28. Miller D. A., Firschein I. L., Dev V. G., Tantravahi R., Miller O. J. The gorilla karyotype: chromosome lengths and polymorphisms. Cytogenet Cell Genet. 1974;13(6):536–550. doi: 10.1159/000130305. [DOI] [PubMed] [Google Scholar]
  29. Myklebost O., Rogne S., Olaisen B., Gedde-Dahl T., Jr, Prydz H. The locus for apolipoprotein CII is closely linked to the apolipoprotein E locus on chromosome 19 in man. Hum Genet. 1984;67(3):309–312. doi: 10.1007/BF00291359. [DOI] [PubMed] [Google Scholar]
  30. Olaisen B., Teisberg P., Gedde-Dahl T., Jr The locus for apolipoprotein E (apoE) is linked to the complement component C3 (C3) locus on chromosome 19 in man. Hum Genet. 1982;62(3):233–236. doi: 10.1007/BF00333526. [DOI] [PubMed] [Google Scholar]
  31. Rees A., Shoulders C. C., Stocks J., Galton D. J., Baralle F. E. DNA polymorphism adjacent to human apoprotein A-1 gene: relation to hypertriglyceridaemia. Lancet. 1983 Feb 26;1(8322):444–446. doi: 10.1016/s0140-6736(83)91440-x. [DOI] [PubMed] [Google Scholar]
  32. Schamaun O., Olaisen B., Mevåg B., Gedde-Dahl T., Jr, Ehnholm C., Teisberg P. The two apolipoprotein loci apo A-I and apo A-IV are closely linked in man. Hum Genet. 1984;68(2):181–184. doi: 10.1007/BF00279311. [DOI] [PubMed] [Google Scholar]
  33. Schumaker V. N., Robinson M. T., Curtiss L. K., Butler R., Sparkes R. S. Anti-apoprotein B monoclonal antibodies detect human low density lipoprotein polymorphism. J Biol Chem. 1984 May 25;259(10):6423–6430. [PubMed] [Google Scholar]
  34. Scott J., Knott T. J., Priestley L. M., Robertson M. E., Mann D. V., Kostner G., Miller G. J., Miller N. E. High-density lipoprotein composition is altered by a common DNA polymorphism adjacent to apoprotein AII gene in man. Lancet. 1985 Apr 6;1(8432):771–773. doi: 10.1016/s0140-6736(85)91443-6. [DOI] [PubMed] [Google Scholar]
  35. Shiloh Y., Shipley J., Brodeur G. M., Bruns G., Korf B., Donlon T., Schreck R. R., Seeger R., Sakai K., Latt S. A. Differential amplification, assembly, and relocation of multiple DNA sequences in human neuroblastomas and neuroblastoma cell lines. Proc Natl Acad Sci U S A. 1985 Jun;82(11):3761–3765. doi: 10.1073/pnas.82.11.3761. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Tata F., Henry I., Markham A. F., Wallis S. C., Weil D., Grzeschik K. H., Junien C., Williamson R., Humphries S. E. Isolation and characterisation of a cDNA clone for human apolipoprotein CI and assignment of the gene to chromosome 19. Hum Genet. 1985;69(4):345–349. doi: 10.1007/BF00291654. [DOI] [PubMed] [Google Scholar]
  37. Wallis S. C., Donald J. A., Forrest L. A., Williamson R., Humphries S. E. The isolation of a genomic clone containing the apolipoprotein CII gene and the detection of linkage disequilibrium between two common DNA polymorphisms around the gene. Hum Genet. 1984;68(4):286–289. doi: 10.1007/BF00292585. [DOI] [PubMed] [Google Scholar]

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