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Expression of Rabbit Globin mRNA Injected into Fused HeLa Cells

  • G. Huez
  • C. Bruck
  • D. Portetelle
  • Y. Cleuter
Part of the NATO Advanced Study Institutes Series book series (NSSA, volume 31)

Abstract

Introduction into a living cell of an isolated messenger RNA allows to answer many interesting questions concerning its translation as well as its stability (see article by Marbaix and Huez). So far the translation of a messenger RNA, upon its manual injection into somatic cells using glass capillaries, has been demonstrated by means of autoradiography (1) or immunofluorescence (2) of the recipient cells. As these detection methods present some limitations, it would be useful in some cases to use classical biochemical methods like gel electrophoresis (followed by autoradiography) to analyse the translation products of an injected mRNA. This implies that a relatively large amount of labelled proteins has to be extracted from the injected cells. This can, of course, be obtained by increasing the number of injected cells. The injection of a large number of cells (over one thousand) is, however, a tedious work. A better approach is to fuse the cells before injection as recently proposed by Graessmann et al. (3). By this way, giant cells are formed which can be injected very easily. The equivalent of one thousand of individual cells can indeed be injected in a matter of minutes.

Keywords

Somatic Cell Giant Cell Translation Product BeLa Cell Manual Injection 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. 1).
    Graessmann, A. and Graessmann, M. (1971). Über die Bildung von Melanin in Muskelzellen nach der Direkten Übertragung von RNA aus Harding-Passay-Melanomzellen. Hoppe-Seyler’s. 2. Physiol. Chem., 352, 527.CrossRefGoogle Scholar
  2. 2).
    Stacey, D.W. and Allfrey, V.G. (1976). Microinjection studies of duck globin messenger RNA translation in human and avian cells. Cell, 9, 725.PubMedCrossRefGoogle Scholar
  3. 3).
    Graessmann, A, Graessmann, M. and Mueller, C (1979). Fused cells are suited for microinjection, Biochem. Biophys. Res. Commun., 88, 428.PubMedCrossRefGoogle Scholar
  4. 4).
    Huez, G. Bruck, C., Portetelle, D. and Cleuter, Y. (1979). Translation of rabbit globin mRNA upon injection into fused HeLa cells. FEBS Lett., in press.Google Scholar
  5. 5).
    Nokin, P., Huez, G. Marbaix, G, Burny, A. and Chantrenne, H. (1976). Molecular modifications associated with aging of globin mRNA in vivo. Eur. J. Biochem., 62, 509.PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1980

Authors and Affiliations

  • G. Huez
    • 1
  • C. Bruck
    • 1
  • D. Portetelle
    • 1
    • 2
  • Y. Cleuter
    • 1
  1. 1.Laboratoire de Chimie Biologique, Département de Biologie MoléculaireUniversité Libre de BruxellesRhode St-GenèseBelgium
  2. 2.Faculté Agronomique de l’EtatGemblouxBelgium

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