Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1982 Jun;79(11):3628–3631. doi: 10.1073/pnas.79.11.3628

Use of restriction endonucleases for mapping the allele for beta s-globin.

J T Wilson, P F Milner, M E Summer, F S Nallaseth, H E Fadel, R H Reindollar, P G McDonough, L B Wilson
PMCID: PMC346476  PMID: 6285354

Abstract

We have reported the direct analysis of the allele for beta 2-globin by using restriction endonuclease Dde I coupled with blot-hybridization analysis. In that report we predicted that a major use of our analysis could be for the prenatal diagnosis of sickle cell anemia. Here we present such an analysis. In addition, this report also describes the use of a new enzyme Mst II, which also distinguish the beta s allele from the normal beta-globin allele. Blot-hybridization analysis with restriction endonuclease Mst II shows the 5' end of the normal beta-globin gene to reside on a fragment of approximately 1.14 kilobases, whereas the 5' end of the beta s-globin gene resides on a fragment of approximately 1.34 kilobases. Because the fragment sizes generated by Mst II are significantly larger than those generated by Dde I, one can easily perform a prenatal diagnosis for sickle cell by standard blot hybridizations onto nitrocellulose filters.

Full text

PDF
3628

Images in this article

3630Image
on p.3630
3630Image
on p.3630
3631Image
on p.3631

Selected References

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

  1. Bittner M., Kupferer P., Morris C. F. Electrophoretic transfer of proteins and nucleic acids from slab gels to diazobenzyloxymethyl cellulose or nitrocellulose sheets. Anal Biochem. 1980 Mar 1;102(2):459–471. doi: 10.1016/0003-2697(80)90182-7. [DOI] [PubMed] [Google Scholar]
  2. Blin N., Stafford D. W. A general method for isolation of high molecular weight DNA from eukaryotes. Nucleic Acids Res. 1976 Sep;3(9):2303–2308. doi: 10.1093/nar/3.9.2303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chang J. C., Kan Y. W. Antenatal diagnosis of sickle cell anaemia by direct analysis of the sickle mutation. Lancet. 1981 Nov 21;2(8256):1127–1129. doi: 10.1016/s0140-6736(81)90584-5. [DOI] [PubMed] [Google Scholar]
  4. Geever R. F., Wilson L. B., Nallaseth F. S., Milner P. F., Bittner M., Wilson J. T. Direct identification of sickle cell anemia by blot hybridization. Proc Natl Acad Sci U S A. 1981 Aug;78(8):5081–5085. doi: 10.1073/pnas.78.8.5081. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Jeffreys A. J. DNA sequence variants in the G gamma-, A gamma-, delta- and beta-globin genes of man. Cell. 1979 Sep;18(1):1–10. doi: 10.1016/0092-8674(79)90348-9. [DOI] [PubMed] [Google Scholar]
  6. Kan Y. W., Dozy A. M. Polymorphism of DNA sequence adjacent to human beta-globin structural gene: relationship to sickle mutation. Proc Natl Acad Sci U S A. 1978 Nov;75(11):5631–5635. doi: 10.1073/pnas.75.11.5631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Kan Y. W., Golbus M. S., Dozy A. M. Prenatal diagnosis of alpha-thalassemia. Clinical application of molecular hybridization. N Engl J Med. 1976 Nov 18;295(21):1165–1167. doi: 10.1056/NEJM197611182952104. [DOI] [PubMed] [Google Scholar]
  8. Levy A., Frei E., Noll M. Efficient transfer of highly resolved small DNA fragments from polyacrylamide gels to DBM paper. Gene. 1980 Nov;11(3-4):283–290. doi: 10.1016/0378-1119(80)90068-2. [DOI] [PubMed] [Google Scholar]
  9. Marotta C. A., Wilson J. T., Forget B. G., Weissman S. M. Human beta-globin messenger RNA. III. Nucleotide sequences derived from complementary DNA. J Biol Chem. 1977 Jul 25;252(14):5040–5053. [PubMed] [Google Scholar]
  10. Orkin S. H., Alter B. P., Altay C., Mahoney M. J., Lazarus H., Hobbins J. C., Nathan D. G. Application of endonuclease mapping to the analysis and prenatal diagnosis of thalassemias caused by globin-gene deletion. N Engl J Med. 1978 Jul 27;299(4):166–172. doi: 10.1056/NEJM197807272990403. [DOI] [PubMed] [Google Scholar]
  11. Phillips J. A., 3rd, Panny S. R., Kazazian H. H., Jr, Boehm C. D., Scott A. F., Smith K. D. Prenatal diagnosis of sickle cell anemia by restriction and endonuclease analysis: HindIII polymorphisms in gamma-globin genes extend test applicability. Proc Natl Acad Sci U S A. 1980 May;77(5):2853–2856. doi: 10.1073/pnas.77.5.2853. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Phillips J. A., 3rd, Scott A. F., Kazazian H. H., Jr, Smith K. D., Stetten G., Thomas G. H. Prenatal diagnosis of hemoglobinopathies by restriction endonuclease analysis: pregnancies at risk for sickle cell anemia and S--O Arab disease. Johns Hopkins Med J. 1979 Aug;145(2):57–60. [PubMed] [Google Scholar]
  13. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  14. Serjeant G. R., Ashcroft M. T., Serjeant B. E., Milner P. F. The clinical features of sickle-cell- thalassaemia in Jamaica. Br J Haematol. 1973 Jan;24(1):19–30. doi: 10.1111/j.1365-2141.1973.tb05723.x. [DOI] [PubMed] [Google Scholar]
  15. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  16. Tuan D., Biro P. A., deRiel J. K., Lazarus H., Forget B. G. Restriction endonuclease mapping of the human gamma globin gene loci. Nucleic Acids Res. 1979 Jun 11;6(7):2519–2544. doi: 10.1093/nar/6.7.2519. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES