Abstract
Normal human metaphase chromosomes isolated from fibroblasts were resolved into 14 peaks based on total Hoechst 33258 fluorescence and sorted with the fluorescence-activated cell sorter. The chromosomal DNA was extracted and characterized by EcoRI analysis. As expected, analysis of the peak containing chromosomes 16 and 18 detected the alpha-globin genes and of the peak containing chromosomes 9, 10, 11, and 12 detected the beta-, gamma-, and delta-globin genes. Translocations were then used to localize further the beta-, gamma-, and delta-globin genes. The first translocation t(11;22)(q25;q11), which moved nearly all of chromosome 11 to a different peak, confirmed that the beta-, gamma-, and delta-globin genes are on this chromosome. The second, t(4;11)(q25;q13), which moved the distal portion of the long arm of chromosome 11 to a new peak, showed that the genes are not in this segment. The third, t(X;11)(q11;p13), moved the distal region of the short arm of chromosome 11 to a peak which now contained the beta-, gamma-, and delta-globin genes. Therefore, the beta-, gamma-, and delta-globin genes residue on the distal portion of the chromosome 11 short arm including bands p13, p14, and p15. This sorting method may be used generally to assign other genes to chromosomal segments of the entire chromosome complement.
Full text
PDFImages in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Anderson W. F., Deisseroth A. B., Velez R., Nienhuis A. W., Ruddle F. H., Kucherlapati R. S. A new technique for mapping the human hemoglobin genes. Cytogenet Cell Genet. 1976;16(1-5):367–371. doi: 10.1159/000130634. [DOI] [PubMed] [Google Scholar]
- Atwood K. C., Henderson A. S., Kacian D., Eicher E. M. On the feasibility of mapping low-multiplicity genes by in situ by hybridization. Cytogenet Cell Genet. 1975;14(3-6):229–231. doi: 10.1159/000130350. [DOI] [PubMed] [Google Scholar]
- BAGLIONI C. The fusion of two peptide chains in hemoglobin Lepore and its interpretation as a genetic deletion. Proc Natl Acad Sci U S A. 1962 Nov 15;48:1880–1886. doi: 10.1073/pnas.48.11.1880. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bishop J. O., Jones K. W. Chromosomal localization of human haemoglobin structural genes. Nature. 1972 Nov 17;240(5377):149–150. doi: 10.1038/240149a0. [DOI] [PubMed] [Google Scholar]
- Carrano A. V., Gray J. W., Langlois R. G., Burkhart-Schultz K. J., Van Dilla M. A. Measurement and purification of human chromosomes by flow cytometry and sorting. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1382–1384. doi: 10.1073/pnas.76.3.1382. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cheung S. W., Tishler P. V., Atkins L., Sengupta S. K., Modest E. J., Forget B. G. Gene mapping by fluorescent in situ hybridization. Cell Biol Int Rep. 1977 May;1(3):255–262. doi: 10.1016/0309-1651(77)90050-9. [DOI] [PubMed] [Google Scholar]
- Deisseroth A., Hendrick D. Human alpha-globin gene expression following chromosomal dependent gene transfer into mouse erythroleukemia cells. Cell. 1978 Sep;15(1):55–63. doi: 10.1016/0092-8674(78)90082-x. [DOI] [PubMed] [Google Scholar]
- Deisseroth A., Nienhuis A., Lawrence J., Giles R., Turner P., Ruddle F. H. Chromosomal localization of human beta globin gene on human chromosome 11 in somatic cell hybrids. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1456–1460. doi: 10.1073/pnas.75.3.1456. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Deisseroth A., Nienhuis A., Turner P., Velez R., Anderson W. F., Ruddle F., Lawrence J., Creagan R., Kucherlapati R. Localization of the human alpha-globin structural gene to chromosome 16 in somatic cell hybrids by molecular hybridization assay. Cell. 1977 Sep;12(1):205–218. doi: 10.1016/0092-8674(77)90198-2. [DOI] [PubMed] [Google Scholar]
- Deisseroth A., Velez R., Nienhuis A. W. Hemoglobin synthesis in somatic cell hybrids: independent segregation of the human alpha- and beta-globin genes. Science. 1976 Mar 26;191(4233):1262–1264. doi: 10.1126/science.943846. [DOI] [PubMed] [Google Scholar]
- Flavell R. A., Kooter J. M., De Boer E., Little P. F., Williamson R. Analysis of the beta-delta-globin gene loci in normal and Hb Lepore DNA: direct determination of gene linkage and intergene distance. Cell. 1978 Sep;15(1):25–41. doi: 10.1016/0092-8674(78)90080-6. [DOI] [PubMed] [Google Scholar]
- Gandini E., Dallapiccola B., Laurent C., Suerinc E. F., Forabosco A., Conconi F., Del Senno L. Evidence for localisation of genes for human alpha-globin on the long arm of chromosome 4. Nature. 1977 Jan 6;265(5589):65–66. doi: 10.1038/265065a0. [DOI] [PubMed] [Google Scholar]
- Goss S. J., Harris H. New method for mapping genes in human chromosomes. Nature. 1975 Jun 26;255(5511):680–684. doi: 10.1038/255680a0. [DOI] [PubMed] [Google Scholar]
- Horan P. K., Wheeless L. L., Jr Quantitative single cell analysis and sorting. Science. 1977 Oct 14;198(4313):149–157. doi: 10.1126/science.905822. [DOI] [PubMed] [Google Scholar]
- Hughes S. H., Stubblefield E., Payvar F., Engel J. D., Dodgson J. B., Spector D., Cordell B., Schimke R. T., Varmus H. E. Gene localization by chromosome fractionation: globin genes are on at least two chromosomes and three estrogen-inducible genes are on three chromosomes. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1348–1352. doi: 10.1073/pnas.76.3.1348. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Huisman T. H., Wrightstone R. N., Wilson J. B., Schroeder W. A., Kendall A. G. Hemoglobin Kenya, the product of fusion of amd polypeptide chains. Arch Biochem Biophys. 1972 Dec;153(2):850–853. doi: 10.1016/0003-9861(72)90408-0. [DOI] [PubMed] [Google Scholar]
- 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]
- Kendall A. G., Ojwang P. J., Schroeder W. A., Huisman T. H. Hemoglobin Kenya, the product of a gamma-beta fusion gene: studies of the family. Am J Hum Genet. 1973 Sep;25(5):548–563. [PMC free article] [PubMed] [Google Scholar]
- Lawn R. M., Fritsch E. F., Parker R. C., Blake G., Maniatis T. The isolation and characterization of linked delta- and beta-globin genes from a cloned library of human DNA. Cell. 1978 Dec;15(4):1157–1174. doi: 10.1016/0092-8674(78)90043-0. [DOI] [PubMed] [Google Scholar]
- Leif R. C., Easter H. N., Jr, Warters R. L., Thomas R. A., Dunlap L. A., Austin M. F. Centrifugal cytology. I. A quantitative technique for the preparation of glutaraldehyde-fixed cells for the light and scanning electron microscope. J Histochem Cytochem. 1971 Apr;19(4):203–215. doi: 10.1177/19.4.203. [DOI] [PubMed] [Google Scholar]
- Little P. F., Flavell R. A., Kooter J. M., Annison G., Williamson R. Structure of the human fetal globin gene locus. Nature. 1979 Mar 15;278(5701):227–231. doi: 10.1038/278227a0. [DOI] [PubMed] [Google Scholar]
- Mears J. G., Ramirez F., Leibowitz D., Bank A. Organization of human delta--and beta-globin genes in cellular DNA and the presence of intragenic inserts. Cell. 1978 Sep;15(1):15–23. doi: 10.1016/0092-8674(78)90079-x. [DOI] [PubMed] [Google Scholar]
- Padgett T. G., Stubbledield E., Varmus H. E. Chicken macrochromosomes contain an endogenous provirus and microchromosomes contain sequences related to the transforming gene of ASV. Cell. 1977 Apr;10(4):649–657. doi: 10.1016/0092-8674(77)90098-8. [DOI] [PubMed] [Google Scholar]
- Prensky W., Holmquist G. Chromosomal localization of human haemoglobin structural genes: techniques queried. Nature. 1973 Jan 5;241(5384):44–45. doi: 10.1038/241044a0. [DOI] [PubMed] [Google Scholar]
- Price P. M., Conover J. H., Hirschhorn K. Chromosomal localization of human haemoglobin structural genes. Nature. 1972 Jun 9;237(5354):340–342. doi: 10.1038/237340a0. [DOI] [PubMed] [Google Scholar]
- Price P. M., Hirschhorn K. In situ hybridization for gene mapping. Birth Defects Orig Artic Ser. 1975;11(3):225–231. [PubMed] [Google Scholar]
- Price P. M., Hirschhorn K. In situ hybridization of chromosome loci. Fed Proc. 1975 Dec;34(13):2227–2232. [PubMed] [Google Scholar]
- SCHWARTZ H. C., SPAET T. H., ZUELZER W. W., NEEL J. V., ROBINSON A. R., KAUFMAN S. F. Combinations of hemoglobin G, hemoglobin S and thalassemia occurring in one family. Blood. 1957 Mar;12(3):238–250. [PubMed] [Google Scholar]
- Weitkamp L. R., Stamatoyannopoulos G., Rowley P. T., Kirk R. L. The linkage relationships of the haemoglobin beta, delta and alpha loci with 34 genetic marker systems. Ann Hum Genet. 1977 Jul;41(1):61–75. doi: 10.1111/j.1469-1809.1977.tb01962.x. [DOI] [PubMed] [Google Scholar]
- Wood W. G. Haemoglobin synthesis during human fetal development. Br Med Bull. 1976 Sep;32(3):282–287. doi: 10.1093/oxfordjournals.bmb.a071376. [DOI] [PubMed] [Google Scholar]