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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
. 1982 Jan;79(1):146–150. doi: 10.1073/pnas.79.1.146

Detection of genes coding for human differentiation markers by their transient expression after DNA transfer.

L J Chang, C L Gamble, C A Izaguirre, M D Minden, T W Mak, E A McCulloch
PMCID: PMC345679  PMID: 6275384

Abstract

We have developed an assay for specific genes in DNA based on transient expression. DNA prepared from patients with acute myeloblastic or acute lymphoblastic leukemia or from the continuous leukemic cell line HL60 was transferred to LTA cells; 48-72 hr later, these recipients expressed hemopoietic differentiation markers as detected by monoclonal antibodies against My-1 (granulocyte specific) and OK-T3 (T-cell specific) using immunofluorescence. The efficiency of transfer was dose and time dependent. We found that genes not expressed in the original cells were expressed after transfer by using this assay. Restriction enzyme analysis showed that My-1 was not expressed with DNA that had been incubated before transfer with either HindIII or Sal I but was present after digestion with either EcoRI or BamHI; after digestion of DNA with the same enzymes, OK-T3 expression was observed only in the HindIII-treated DNA. These studies indicate that DNA was transferred; this approach may provide an efficient method for detecting the activity of specific genes in the absence of selection.

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Selected References

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  1. Capecchi M. R. High efficiency transformation by direct microinjection of DNA into cultured mammalian cells. Cell. 1980 Nov;22(2 Pt 2):479–488. doi: 10.1016/0092-8674(80)90358-x. [DOI] [PubMed] [Google Scholar]
  2. Civin C. I., Mirro J., Banquerigo M. L. My-1, new myeloid-specific antigen identified by a mouse monoclonal antibody. Blood. 1981 May;57(5):842–845. [PubMed] [Google Scholar]
  3. Collins S. J., Gallo R. C., Gallagher R. E. Continuous growth and differentiation of human myeloid leukaemic cells in suspension culture. Nature. 1977 Nov 24;270(5635):347–349. doi: 10.1038/270347a0. [DOI] [PubMed] [Google Scholar]
  4. Cooper G. M., Okenquist S., Silverman L. Transforming activity of DNA of chemically transformed and normal cells. Nature. 1980 Apr 3;284(5755):418–421. doi: 10.1038/284418a0. [DOI] [PubMed] [Google Scholar]
  5. Flavell R. A., Bernards R., Kooter J. M., de Boer E., Little P. F., Annison G., Williamson R. The structure of the human beta-globin gene in beta-thalassaemia. Nucleic Acids Res. 1979 Jun 25;6(8):2749–2760. doi: 10.1093/nar/6.8.2749. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fritsch E. F., Lawn R. M., Maniatis T. Characterisation of deletions which affect the expression of fetal globin genes in man. Nature. 1979 Jun 14;279(5714):598–603. doi: 10.1038/279598a0. [DOI] [PubMed] [Google Scholar]
  7. Graham F. L., van der Eb A. J. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology. 1973 Apr;52(2):456–467. doi: 10.1016/0042-6822(73)90341-3. [DOI] [PubMed] [Google Scholar]
  8. Jeffreys A. J. DNA sequence variants in the G gamma-, A gamma-, delta- and beta-globin genes of man. Cell. 1979 Sep;18(1):1–10. doi: 10.1016/0092-8674(79)90348-9. [DOI] [PubMed] [Google Scholar]
  9. Krontiris T. G., Cooper G. M. Transforming activity of human tumor DNAs. Proc Natl Acad Sci U S A. 1981 Feb;78(2):1181–1184. doi: 10.1073/pnas.78.2.1181. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kung P., Goldstein G., Reinherz E. L., Schlossman S. F. Monoclonal antibodies defining distinctive human T cell surface antigens. Science. 1979 Oct 19;206(4416):347–349. doi: 10.1126/science.314668. [DOI] [PubMed] [Google Scholar]
  11. Milman G., Herzberg M. Efficient DNA transfection and rapid assay for thymidine kinase activity and viral antigenic determinants. Somatic Cell Genet. 1981 Mar;7(2):161–170. doi: 10.1007/BF01567655. [DOI] [PubMed] [Google Scholar]
  12. Minden M. D., Buick R. N., McCulloch E. A. Separation of blast cell and T-lymphocyte progenitors in the blood of patients with acute myeloblastic leukemia. Blood. 1979 Jul;54(1):186–195. [PubMed] [Google Scholar]
  13. Mulligan R. C., Berg P. Expression of a bacterial gene in mammalian cells. Science. 1980 Sep 19;209(4463):1422–1427. doi: 10.1126/science.6251549. [DOI] [PubMed] [Google Scholar]
  14. Pellicer A., Robins D., Wold B., Sweet R., Jackson J., Lowy I., Roberts J. M., Sim G. K., Silverstein S., Axel R. Altering genotype and phenotype by DNA-mediated gene transfer. Science. 1980 Sep 19;209(4463):1414–1422. doi: 10.1126/science.7414320. [DOI] [PubMed] [Google Scholar]
  15. Sharp P. A., Sugden B., Sambrook J. Detection of two restriction endonuclease activities in Haemophilus parainfluenzae using analytical agarose--ethidium bromide electrophoresis. Biochemistry. 1973 Jul 31;12(16):3055–3063. doi: 10.1021/bi00740a018. [DOI] [PubMed] [Google Scholar]
  16. Shih C., Shilo B. Z., Goldfarb M. P., Dannenberg A., Weinberg R. A. Passage of phenotypes of chemically transformed cells via transfection of DNA and chromatin. Proc Natl Acad Sci U S A. 1979 Nov;76(11):5714–5718. doi: 10.1073/pnas.76.11.5714. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Shilo B. Z., Weinberg R. A. Unique transforming gene in carcinogen-transformed mouse cells. Nature. 1981 Feb 12;289(5798):607–609. doi: 10.1038/289607a0. [DOI] [PubMed] [Google Scholar]
  18. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  19. Wigler M., Pellicer A., Silverstein S., Axel R., Urlaub G., Chasin L. DNA-mediated transfer of the adenine phosphoribosyltransferase locus into mammalian cells. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1373–1376. doi: 10.1073/pnas.76.3.1373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Wigler M., Sweet R., Sim G. K., Wold B., Pellicer A., Lacy E., Maniatis T., Silverstein S., Axel R. Transformation of mammalian cells with genes from procaryotes and eucaryotes. Cell. 1979 Apr;16(4):777–785. doi: 10.1016/0092-8674(79)90093-x. [DOI] [PubMed] [Google Scholar]

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