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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
. 1975 May;72(5):1868–1872. doi: 10.1073/pnas.72.5.1868

Identification of human RNA transcripts among heterogeneous nuclear RNA from man-mouse somatic cell hybrids.

R A Di Cioccio, R Voss, M Krim, K H Grzeschik, M Siniscalco
PMCID: PMC432648  PMID: 1057177

Abstract

In man-mouse hybrid line from our cell library, the only cytological detectable portion of the human genome is the X chromosome, and the only genetic markers regularly expressed are coded by genes known to be ?X-linked. A component of the heterogeneous nuclear RNA of these cells was found to be complementary to repetitive human DNA sequences by means of RNA-DNA hybridization on nitrocellulose filters. The same procedure also permitted the identification of hybrid cell DNA sequences that are complementary to human heterogeneous nuclear RNA. This experimental approach, coupled with hybridization studies in situ, is expected to yield critical data on the distribution and the specificity of the repetitive DNA sequences present in the human genome and to provide a new tool for cytological mapping of human chromosomes.

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Selected References

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  1. Arrighi F. E., Hsu T. C. Localization of heterochromatin in human chromosomes. Cytogenetics. 1971;10(2):81–86. doi: 10.1159/000130130. [DOI] [PubMed] [Google Scholar]
  2. Caspersson T., Zech L., Johansson C., Modest E. J. Identification of human chromosomes by DNA-binding fluorescent agents. Chromosoma. 1970;30(2):215–227. doi: 10.1007/BF00282002. [DOI] [PubMed] [Google Scholar]
  3. Coon H. G., Horak I., Dawid I. B. Propagation of both parental mitochondrial DNAs in rat-human and mouse-human hybrid cells. J Mol Biol. 1973 Dec 15;81(3):285–298. doi: 10.1016/0022-2836(73)90142-3. [DOI] [PubMed] [Google Scholar]
  4. Cox R. P., Krauss M. R., Balis M. E., Dancis J. Communication between normal and enzyme deficient cells in tissue culture. Exp Cell Res. 1972 Sep;74(1):251–268. doi: 10.1016/0014-4827(72)90503-4. [DOI] [PubMed] [Google Scholar]
  5. Croce C. M., Litwack G., Koprowski H. Human regulatory gene for inducible tyrosine aminotransferase in rat-human hybrids. Proc Natl Acad Sci U S A. 1973 Apr;70(4):1268–1272. doi: 10.1073/pnas.70.4.1268. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gillespie D., Spiegelman S. A quantitative assay for DNA-RNA hybrids with DNA immobilized on a membrane. J Mol Biol. 1965 Jul;12(3):829–842. doi: 10.1016/s0022-2836(65)80331-x. [DOI] [PubMed] [Google Scholar]
  7. Grzeschik K. H., Allderdice P. W., Grzeschik A., Opitz J. M., Miller O. J., Siniscalco M. Cytological mapping of human X-linked genes by use of somatic cell hybrids involving an X-autosome translocation (mouse-hamster-human X-linked markers). Proc Natl Acad Sci U S A. 1972 Jan;69(1):69–73. doi: 10.1073/pnas.69.1.69. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kumar A., Lindberg U. Characterization of messenger ribonucleoprotein and messenger RNA from KB cells. Proc Natl Acad Sci U S A. 1972 Mar;69(3):681–685. doi: 10.1073/pnas.69.3.681. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Káo F. T., Puck T. T. Genetics of somatic mammalian cells: linkage studies with human-Chinese hamster cell hybrids. Nature. 1970 Oct 24;228(5269):329–332. doi: 10.1038/228329a0. [DOI] [PubMed] [Google Scholar]
  10. LITTLEFIELD J. W. THREE DEGREES OF GUANYLIC ACID--INOSINIC ACID PYROPHOSPHORYLASE DEFICIENCY IN MOUSE FIBROBLASTS. Nature. 1964 Sep 12;203:1142–1144. doi: 10.1038/2031142a0. [DOI] [PubMed] [Google Scholar]
  11. Levine E. M. Mycoplasma contamination of animal cell cultures: a simple, rapid detection method. Exp Cell Res. 1972 Sep;74(1):99–109. doi: 10.1016/0014-4827(72)90484-3. [DOI] [PubMed] [Google Scholar]
  12. Littlefield J. W. The use of drug-resistant markers to study the hybridization of mouse fibroblasts. Exp Cell Res. 1966 Jan;41(1):190–196. doi: 10.1016/0014-4827(66)90558-1. [DOI] [PubMed] [Google Scholar]
  13. Meera Khan P. Enzyme electrophoresis on cellulose acetate gel: zymogram patterns in mgh-mouse and man--Chinese hamster somatic cell hybrids. Arch Biochem Biophys. 1971 Aug;145(2):470–483. doi: 10.1016/s0003-9861(71)80007-3. [DOI] [PubMed] [Google Scholar]
  14. Meera Khan P., Westerveld A., Grzeschik K. H., Deys B. F., Garson O. M., Siniscalco M. X-linkage of human phosphoglycerate kinase confirmed in man-mouse and man-Chinese hamster somatic cell hybrids. Am J Hum Genet. 1971 Nov;23(6):614–623. [PMC free article] [PubMed] [Google Scholar]
  15. Migeon B. R., Smith S. W., Leddy C. L. The nature of thymidine kinase in the human-mouse hybrid cell. Biochem Genet. 1969 Dec;3(6):583–590. doi: 10.1007/BF00485479. [DOI] [PubMed] [Google Scholar]
  16. Miller O. J., Cook P. R., Meera Khan P., Shin S., Siniscalco M. Mitotic separation of two human X-linked genes in man--mouse somatic cell hybrids. Proc Natl Acad Sci U S A. 1971 Jan;68(1):116–120. doi: 10.1073/pnas.68.1.116. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Miller O. J., Miller D. A., Allderdice P. W., Dev V. G., Grewal M. S. Quinacrine fluorescent karyotypes of human diploid and heteroploid cell lines. Cytogenetics. 1971;10(5):338–346. doi: 10.1159/000130152. [DOI] [PubMed] [Google Scholar]
  18. Penman S. RNA metabolism in the HeLa cell nucleus. J Mol Biol. 1966 May;17(1):117–130. doi: 10.1016/s0022-2836(66)80098-0. [DOI] [PubMed] [Google Scholar]
  19. SZYBALSKI W., SZYBALSKA E. H. Drug sensitivity as a genetic marker for human cell lines. Med Bull (Ann Arbor) 1962 Sep-Oct;28:277–293. [PubMed] [Google Scholar]
  20. Seabright M. The use of proteolytic enzymes for the mapping of structural rearrangements in the chromosomes of man. Chromosoma. 1972;36(2):204–210. doi: 10.1007/BF00285214. [DOI] [PubMed] [Google Scholar]
  21. Seegmiller J. E., Rosenbloom F. M., Kelley W. N. Enzyme defect associated with a sex-linked human neurological disorder and excessive purine synthesis. Science. 1967 Mar 31;155(3770):1682–1684. doi: 10.1126/science.155.3770.1682. [DOI] [PubMed] [Google Scholar]
  22. Soeiro R., Darnell J. E. Competition hybridization by "pre-saturation" of HeLa cell DNA. J Mol Biol. 1969 Sep 28;44(3):551–562. doi: 10.1016/0022-2836(69)90379-9. [DOI] [PubMed] [Google Scholar]
  23. Weiss M. C., Green H. Human-mouse hybrid cell lines containing partial complements of human chromosomes and functioning human genes. Proc Natl Acad Sci U S A. 1967 Sep;58(3):1104–1111. doi: 10.1073/pnas.58.3.1104. [DOI] [PMC free article] [PubMed] [Google Scholar]

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