Skip to main content
Log in

Assignment of snRNA gene sequences to the large chromosomes of rat kangaroo and Chinese hamster isolated by flow cytometric sorting

  • Published:
Chromosoma Aims and scope Submit manuscript

Abstract

Chromosomes from a rat kangaroo (Potorous tridactylus) cell line (PtK2) and from a Chinese hamster (Cricetulus griseus) cell line (CHV79) were isolated by means of fluorescence activated flow cytometric sorting. DAPI (4′-6-diamino-2-phenylindole) was used as the DNA specific fluorescent dye. The karyotype of the PtK2 cells which exhibits 13 chromosomes was separated into 6, and the 22 chromosomes of the CHV79 cells were resolved into 11 fractions. DNA extracted from these chromosomal fractions was used for restriction enzyme digestion and blotting on nitrocellulose filters. The blots were challenged with gene probes corresponding to ribosomal RNA (18S and 28S) and small nuclear RNA (U1-snRNA) genes. The rRNA genes were exclusively assigned to chromosomes containing the nucleolus organizing region (in PtK2: X chromosome; in CHV79: chromosomes 4, 5, 6, and 11). — Solely the largest chromosomes in both cell lines hybridized with U1-snRNA indicating that these gene sequences are located on those chromosomes only. Further possible genetic and biochemical applications of this experimental system are discussed.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Alonso, A., Krieg, L., Winter, H., Sekeris, C.E.: The synthesis of a cDNA copy complementary to two snRNAs and the localization of their genes in the rat genome. Biochem. biophys. Res. Comm. 95, 148–155 (1980)

    Google Scholar 

  • Alonso, A., Winter, H., Krieg, L.: The large hnRNP complexes of rat liver: Protein and RNA moieties. In: Int. cell biology 1980–1981 (H.G. Schweiger, ed.), pp. 72–76. Berlin-Heidelberg: Springer Verlag 1981

    Google Scholar 

  • Blin, N., Stafford, D.W.: A general method for isolation of high molecular weight DNA from eukaryotes. Nucleic Acids Res. 3, 2303–2308 (1976)

    Google Scholar 

  • Carrano, A.V., Gray, J.W., Moore II, D.H., Minkler, J.L., Mayall, B.H., VanDilla, M.A., Mendelsohn, M.L.: Purification of the chromosomes of the Indian muntjak by flow sorting. J. Histochem. Cytochem. 24, 348–354 (1976)

    Google Scholar 

  • Carrano, A.V., Gray, J.W., Langlois, R.G., Burkhart-Schultz, K.J., VanDilla, M.A.: Measurement and purification of human chromosomes by flow cytometry and sorting. Proc. nat. Acad. Sci. (Wash.) 76, 1382–1384 (1979)

    Google Scholar 

  • Deisseroth, A., Nienhuis, A., Turner, P., Velez, R., Anderson, W.F., Ruddle, F., Lawrence, J., Creagan, R., Kucherlapati, R.: Localization by the human α-globin structural gene to chromosome 16 in somatic cell hybrids by molecular hybridization assay. Cell 12, 205–218 (1977)

    Google Scholar 

  • Denison, A.A., VanArsdell, S.W., Bernstein, L.B., Weiner, A.M.: Abundant pseudogenes for small nuclear RNAs are dispersed in the human genome. Proc. nat. Acad. Sci. (Wash.) 78, 810–814 (1981)

    Google Scholar 

  • Eliceiri, G.L.: Sensitivity of low molecular weight RNA synthesis to UV radiation. Nature (Lond.) 279, 80–81 (1979)

    Google Scholar 

  • Engberg, J., Hellung-Larsen, P., Frederiksen, S.: Isolation and DNA-RNA hybridization properties of small-molecular-weight nuclear RNA components from baby-hamster-kidney cells. Europ. J. Biochem. 41, 321–328 (1974)

    Google Scholar 

  • Flavell, R.A., Kooter, J.M., DeBoer, E., Little, P.F.R., Williamson, R.: Analysis of the β-γ-globin gene loci in normal and Hb lepore DNA: Direct determination of gene linkage and intergene distance. Cell 15, 25–41 (1978)

    Google Scholar 

  • Goodpasture, C., Bloom, S.E.: Visualization of nucleolar organizer regions in mammalian chromosomes using silver staining. Chromosoma (Berl.) 53, 37–50 (1975)

    Google Scholar 

  • Gray, J.W., Carrano, A.V., Steinmetz, C.C., VanDilla, M.A., Moore II, D.H., Mayall, B.H., Mendelssohn, M.L.: Chromosome measurement and sorting by flow systems. Proc. nat. Acad. Sci. (Wash.) 72, 1231–1234 (1975)

    Google Scholar 

  • Gray, J.W., Langlois, R.G., Carrano, A.V., Burkhart-Schultz, K., VanDilla, M.A.: High resolution chromosome analysis: One and two parameter flow cytometry. Chromosoma (Berl.) 73, 9–27 (1979)

    Google Scholar 

  • Grummt, I., Gross, H.J.: Structural organization of mouse rDNA: Comparison of transcribed and non-transcribed regions. Molec. Gen. Genet. 177, 223–229 (1980)

    Google Scholar 

  • Hayashi, K.: Organization of sequences related to UG RNA in the human genome. Nucl. Acids. Res. 3, 3379–3388 (1981)

    Google Scholar 

  • Houck, C.M., Rinehart, F.P., Schmid, C.W.: A ubiquitous family of repeated DNA sequences in the human genome. J. molec. Biol. 132, 289–306 (1979)

    Google Scholar 

  • Hsu, T.C., Spirito, S.E., Pardue, M.L.: Distribution of 18 + 28S ribosomal genes in mammalian genomes. Chromosoma (Berl.) 53, 25–36 (1975)

    Google Scholar 

  • Jones, C.W., Kafatos, F.C.: Coordinately expressed members of two chorion multi-gene families are clustered, alternating and divergently oriented. Nature (Lond.) 284, 635–638 (1980)

    Google Scholar 

  • Kafatos, F., Jones, C.W., Efstratiadis, A.: Determination of nucleic acid sequence homology and relative concentrations by a dot hybridization procedure. Nucleic Acids Res. 7, 1541–1552 (1979)

    Google Scholar 

  • Krayev, A.S., Kramerov, D.A., Skryabin, K.G., Ryskos, A.P., Bayev, A.A., Georgiev, G.P.: The nucleotide sequence of the ubiquitous repetitive DNA sequence B1 complementary to the most abundant class of mouse fold back RNA. Nucleic Acids Res. 8, 1201–1215 (1980)

    Google Scholar 

  • Krol, A., Gallinaro, H., Lazar, E., Jacob, M., Branlant, C.: U5 RNAs are encoded by multiple genes. Nucleic Acids Res. 3, 769–787 (1981)

    Google Scholar 

  • Lawn, R.M., Fritsch, E.F., Parker, R.C., Blake, G., Maniatis, T.: The isolation and characterization of linked β- and γ-globin genes from a cloned library of human DNA. Cell 15, 1157–1175 (1978)

    Google Scholar 

  • Lebo, R.V., Carrano, A.V., Burkhart-Schultz, K., Dozy, A.M., Yu, L.C., Kan, Y.W.: Assignment of human β-, γ, and δ-globin genes to the short arm of chromosome 11 by chromosome sorting and DNA restriction enzyme analysis. Proc. nat. Acad. Sci. (Wash.) 76, 5804–5808 (1979)

    Google Scholar 

  • Manser, T., Gesteland, R.F.: Characterization of small nuclear RNA UI gene. J. molec. appl. Genet. 1, 117–125 (1981)

    Google Scholar 

  • Murray, V., Holliday, R.: Mechanism for RNA splicing of gene transcripts. FEBS Lett. 106, 5–7 (1979)

    Google Scholar 

  • Owerbach, D., Bell, G.I., Rutter, W.J., Shows, T.B.: The insulin gene is located on chromosome 11 in humans. Nature (Lond.) 286, 82–84 (1980)

    Google Scholar 

  • Padgett, T.G., Stubblefield, E., Varmus, H.E.: Chicken macrochromosomes contain an endogenous provirus and microchromosomes contain sequences related to the transforming gene of ASV. Cell 10, 649–657 (1977)

    Google Scholar 

  • Rogers, J., Wall, R.: A mechanism for RNA splicing. Proc. nat. Acad. Sci. (Wash.) 77, 1877–1879 (1980)

    Google Scholar 

  • Royal, A., Garapin, A., Cami, B., Perrin, F., Mandel, J.L., LeMeur, M., Bregegegre, F., Gannon, F., LePennec, J.P., Chambon, P., Kourilsky, P.: The ovalbumin gene region: Common features in the organization of three genes expressed in chicken oviduct under hormonal control. Nature (Lond.) 279, 125–132 (1979)

    Google Scholar 

  • Shaw, M.W., Krooth, R.S.: The chromosomes of the Tasmanian rat-kangaroo. Cytogenetics 3, 19–33 (1964)

    Google Scholar 

  • Scalenghe, F., Turco, E., Edström, J.E., Pirrotta, V., Melli, M.: Microdissection and cloning of DNA from a specific region of Drosophila melanogaster polytene chromosomes. Chromosoma (Berl.) 82, 205–216 (1981)

    Google Scholar 

  • Sillar, R., Young, B.D.: A new method for the preparation of metaphase chromosomes for flow analysis. J. Histochem. Cytochem. 29, 74–78 (1981)

    Google Scholar 

  • Stöhr, M., Hutter, K.J., Frank, M., Futterman, G., Goerttler, K.: A flow cytometric study of chromosomes from rat kangaroo and Chinese hamster cells. Histochemistry 67, 179–190 (1980)

    Google Scholar 

  • Stöhr, M., Hutter, K.J., Frank, M., Goerttler, K.: A reliable preparation of mono-dispersed chromosome suspensions for flow cytometry. Histochemistry, in press (1982)

  • Stubblefield, E., Deavan, L., Cram, L.S.: Flow microfluorometric analysis of isolated Chinese hamster chromosomes. Exp. Cell Res. 94, 464–468 (1975)

    Google Scholar 

  • Wise, J.A., Weiner, A.: Dictyostelium small nuclear RNA D2 is homologous to rat nucleolar RNA U3 and is encoded by a dispersed multigene family. Cell 22, 109–118 (1980)

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Blin, N., Stöhr, M., Hutter, K.J. et al. Assignment of snRNA gene sequences to the large chromosomes of rat kangaroo and Chinese hamster isolated by flow cytometric sorting. Chromosoma 85, 723–733 (1982). https://doi.org/10.1007/BF00330784

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00330784

Keywords

Navigation