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. 1988 Sep;7(9):2801–2805. doi: 10.1002/j.1460-2075.1988.tb03135.x

Sensitive and high resolution in situ hybridization to human chromosomes using biotin labelled probes: assignment of the human thymocyte CD1 antigen genes to chromosome 1.

D G Albertson 1, R Fishpool 1, P Sherrington 1, E Nacheva 1, C Milstein 1
PMCID: PMC457071  PMID: 3053166

Abstract

A method for in situ hybridization originally developed for mapping genes in the nematode, Caenorhabditis elegans has been adapted for high resolution cytological mapping of genes in the human. The probe DNAs are labelled by incorporation of biotin dUTP and the site of hybridization detected by immunofluorescence. For the accurate assignment of the hybridization signal to chromosome bands, visualized by staining with Hoechst 33258, a heterologous ribosomal DNA probe is also included in the hybridization reaction. These rDNA signals are used as fiducial markers when aligning the two fluorescent images. We demonstrate the method by assignment of the human thymocyte CD1 antigen genes to human chromosome 1q22-23.

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Selected References

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  1. Albertson D. G. Localization of the ribosomal genes in Caenorhabditis elegans chromosomes by in situ hybridization using biotin-labeled probes. EMBO J. 1984 Jun;3(6):1227–1234. doi: 10.1002/j.1460-2075.1984.tb01957.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Albertson D. G. Mapping muscle protein genes by in situ hybridization using biotin-labeled probes. EMBO J. 1985 Oct;4(10):2493–2498. doi: 10.1002/j.1460-2075.1985.tb03961.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Calabi F., Milstein C. A novel family of human major histocompatibility complex-related genes not mapping to chromosome 6. Nature. 1986 Oct 9;323(6088):540–543. doi: 10.1038/323540a0. [DOI] [PubMed] [Google Scholar]
  4. Files J. G., Hirsh D. Ribosomal DNA of Caenorhabditis elegans. J Mol Biol. 1981 Jun 25;149(2):223–240. doi: 10.1016/0022-2836(81)90299-0. [DOI] [PubMed] [Google Scholar]
  5. Gall J. G., Pardue M. L. Formation and detection of RNA-DNA hybrid molecules in cytological preparations. Proc Natl Acad Sci U S A. 1969 Jun;63(2):378–383. doi: 10.1073/pnas.63.2.378. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Garson J. A., van den Berghe J. A., Kemshead J. T. Novel non-isotopic in situ hybridization technique detects small (1 Kb) unique sequences in routinely G-banded human chromosomes: fine mapping of N-myc and beta-NGF genes. Nucleic Acids Res. 1987 Jun 25;15(12):4761–4770. doi: 10.1093/nar/15.12.4761. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Langer-Safer P. R., Levine M., Ward D. C. Immunological method for mapping genes on Drosophila polytene chromosomes. Proc Natl Acad Sci U S A. 1982 Jul;79(14):4381–4385. doi: 10.1073/pnas.79.14.4381. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Langer P. R., Waldrop A. A., Ward D. C. Enzymatic synthesis of biotin-labeled polynucleotides: novel nucleic acid affinity probes. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6633–6637. doi: 10.1073/pnas.78.11.6633. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Lawrence J. B., Villnave C. A., Singer R. H. Sensitive, high-resolution chromatin and chromosome mapping in situ: presence and orientation of two closely integrated copies of EBV in a lymphoma line. Cell. 1988 Jan 15;52(1):51–61. doi: 10.1016/0092-8674(88)90530-2. [DOI] [PubMed] [Google Scholar]
  10. Martin L. H., Calabi F., Lefebvre F. A., Bilsland C. A., Milstein C. Structure and expression of the human thymocyte antigens CD1a, CD1b, and CD1c. Proc Natl Acad Sci U S A. 1987 Dec;84(24):9189–9193. doi: 10.1073/pnas.84.24.9189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Martin L. H., Calabi F., Milstein C. Isolation of CD1 genes: a family of major histocompatibility complex-related differentiation antigens. Proc Natl Acad Sci U S A. 1986 Dec;83(23):9154–9158. doi: 10.1073/pnas.83.23.9154. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Mitchell A. R., Ambros P., Gosden J. R., Morten J. E., Porteous D. J. Gene mapping and physical arrangements of human chromatin in transformed, hybrid cells: fluorescent and autoradiographic in situ hybridization compared. Somat Cell Mol Genet. 1986 Jul;12(4):313–324. doi: 10.1007/BF01570725. [DOI] [PubMed] [Google Scholar]
  13. Yunis J. J. Mid-prophase human chromosomes. The attainment of 2000 bands. Hum Genet. 1981;56(3):293–298. doi: 10.1007/BF00274682. [DOI] [PubMed] [Google Scholar]
  14. Yunis J. J., Sawyer J. R., Ball D. W. Characterization of banding patterns of metaphase-prophase G-banded chromosomes and their use in gene mapping. Cytogenet Cell Genet. 1978;22(1-6):679–683. doi: 10.1159/000131052. [DOI] [PubMed] [Google Scholar]

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