Skip to main content
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1987 Jun 25;15(12):4761–4770. doi: 10.1093/nar/15.12.4761

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.

J A Garson, J A van den Berghe, J T Kemshead
PMCID: PMC305916  PMID: 3299258

Abstract

A novel in situ hybridization technique is described. This non-radioactive technique combines, for the first time, the high spacial resolution and rapid signal development of the non-isotopic approach with the previously unrivalled sensitivity of autoradiography. The procedure, which employs biotin labelled DNA probes and a streptavidin-alkaline phosphatase based detection system, is compatible with pre G-banding and can be performed on archival material. Unique sequences as small as 1 Kb are detectable. Using this technique, we have mapped the N-myc oncogene and the gene for beta-Nerve Growth Factor to 2p24 and 1p13 respectively.

Full text

PDF
4761

Images in this article

Selected References

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

  1. Bauman J. G., Wiegant J., Van Duijn P., Lubsen N. H., Sondermeijer P. J., Hennig W., Kubli E. Rapid and high resolution detection of in situ hybridisation to polytene chromosomes using fluorochrome-labeled RNA. Chromosoma. 1981;84(1):1–18. doi: 10.1007/BF00293359. [DOI] [PubMed] [Google Scholar]
  2. Burns J., Chan V. T., Jonasson J. A., Fleming K. A., Taylor S., McGee J. O. Sensitive system for visualising biotinylated DNA probes hybridised in situ: rapid sex determination of intact cells. J Clin Pathol. 1985 Oct;38(10):1085–1092. doi: 10.1136/jcp.38.10.1085. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chandler M. E., Yunis J. J. A high resolution in situ hybridization technique for the direct visualization of labeled G-banded early metaphase and prophase chromosomes. Cytogenet Cell Genet. 1978;22(1-6):352–356. doi: 10.1159/000130970. [DOI] [PubMed] [Google Scholar]
  4. Darby J. K., Feder J., Selby M., Riccardi V., Ferrell R., Siao D., Goslin K., Rutter W., Shooter E. M., Cavalli-Sforza L. L. A discordant sibship analysis between beta-NGF and neurofibromatosis. Am J Hum Genet. 1985 Jan;37(1):52–59. [PMC free article] [PubMed] [Google Scholar]
  5. Francke U., de Martinville B., Coussens L., Ullrich A. The human gene for the beta subunit of nerve growth factor is located on the proximal short arm of chromosome 1. Science. 1983 Dec 16;222(4629):1248–1251. doi: 10.1126/science.6648531. [DOI] [PubMed] [Google Scholar]
  6. Heim S., Mitelman F. Proliferation-specific and differentiation-associated chromosomal breakpoints in human neoplasia--a unifying model. Hereditas. 1986;104(2):307–312. doi: 10.1111/j.1601-5223.1986.tb00544.x. [DOI] [PubMed] [Google Scholar]
  7. Hopman A. H., Wiegant J., van Duijn P. A new hybridocytochemical method based on mercurated nucleic acid probes and sulfhydryl-hapten ligands. II. Effects of variations in ligand structure on the in situ detection of mercurated probes. Histochemistry. 1986;84(2):179–185. doi: 10.1007/BF00499830. [DOI] [PubMed] [Google Scholar]
  8. Jhanwar S. C., Neel B. G., Hayward W. S., Chaganti R. S. Localization of c-ras oncogene family on human germ-line chromosomes. Proc Natl Acad Sci U S A. 1983 Aug;80(15):4794–4797. doi: 10.1073/pnas.80.15.4794. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Landegent J. E., Jansen in de Wal N., Fisser-Groen Y. M., Bakker E., van der Ploeg M., Pearson P. L. Fine mapping of the Huntington disease linked D4S10 locus by non-radioactive in situ hybridization. Hum Genet. 1986 Aug;73(4):354–357. doi: 10.1007/BF00279100. [DOI] [PubMed] [Google Scholar]
  10. Landegent J. E., Jansen in de Wal N., van Ommen G. J., Baas F., de Vijlder J. J., van Duijn P., Van der Ploeg M. Chromosomal localization of a unique gene by non-autoradiographic in situ hybridization. Nature. 1985 Sep 12;317(6033):175–177. doi: 10.1038/317175a0. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Lau Y. F. Detection of Y-specific repeat sequences in normal and variant human chromosomes using in situ hybridization with biotinylated probes. Cytogenet Cell Genet. 1985;39(3):184–187. doi: 10.1159/000132132. [DOI] [PubMed] [Google Scholar]
  13. Leary J. J., Brigati D. J., Ward D. C. Rapid and sensitive colorimetric method for visualizing biotin-labeled DNA probes hybridized to DNA or RNA immobilized on nitrocellulose: Bio-blots. Proc Natl Acad Sci U S A. 1983 Jul;80(13):4045–4049. doi: 10.1073/pnas.80.13.4045. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Malcolm S., Barton P., Murphy C., Ferguson-Smith M. A. Chromosomal localization of a single copy gene by in situ hybridization--human beta globin genes on the short arm of chromosome 11. Ann Hum Genet. 1981 May;45(Pt 2):135–141. doi: 10.1111/j.1469-1809.1981.tb00315.x. [DOI] [PubMed] [Google Scholar]
  15. Manuelidis L., Langer-Safer P. R., Ward D. C. High-resolution mapping of satellite DNA using biotin-labeled DNA probes. J Cell Biol. 1982 Nov;95(2 Pt 1):619–625. doi: 10.1083/jcb.95.2.619. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. McMaster G. K., Carmichael G. G. Analysis of single- and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4835–4838. doi: 10.1073/pnas.74.11.4835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Münke M., Lindgren V., de Martinville B., Francke U. Comparative analysis of mouse-human hybrids with rearranged chromosomes 1 by in situ hybridization and Southern blotting: high-resolution mapping of NRAS, NGFB, and AMY on human chromosome 1. Somat Cell Mol Genet. 1984 Nov;10(6):589–599. doi: 10.1007/BF01535224. [DOI] [PubMed] [Google Scholar]
  18. Ponder B. A., Wilkinson M. M. Inhibition of endogenous tissue alkaline phosphatase with the use of alkaline phosphatase conjugates in immunohistochemistry. J Histochem Cytochem. 1981 Aug;29(8):981–984. doi: 10.1177/29.8.7024402. [DOI] [PubMed] [Google Scholar]
  19. Schwab M., Alitalo K., Klempnauer K. H., Varmus H. E., Bishop J. M., Gilbert F., Brodeur G., Goldstein M., Trent J. Amplified DNA with limited homology to myc cellular oncogene is shared by human neuroblastoma cell lines and a neuroblastoma tumour. Nature. 1983 Sep 15;305(5931):245–248. doi: 10.1038/305245a0. [DOI] [PubMed] [Google Scholar]
  20. Schwab M., Varmus H. E., Bishop J. M., Grzeschik K. H., Naylor S. L., Sakaguchi A. Y., Brodeur G., Trent J. Chromosome localization in normal human cells and neuroblastomas of a gene related to c-myc. Nature. 1984 Mar 15;308(5956):288–291. doi: 10.1038/308288a0. [DOI] [PubMed] [Google Scholar]
  21. Viegas-Péquignot E., Malfoy B., Leng M., Dutrillaux B., Tchen P. In situ hybridization of an acetylaminofluorene-modified probe recognized by Z-DNA antibodies in vitro. Cytogenet Cell Genet. 1986;42(1-2):105–109. doi: 10.1159/000132260. [DOI] [PubMed] [Google Scholar]
  22. Zabel B. U., Eddy R. L., Lalley P. A., Scott J., Bell G. I., Shows T. B. Chromosomal locations of the human and mouse genes for precursors of epidermal growth factor and the beta subunit of nerve growth factor. Proc Natl Acad Sci U S A. 1985 Jan;82(2):469–473. doi: 10.1073/pnas.82.2.469. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES