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Human globin gene expression in hybrid 2S MEL × human fibroblast cells

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Somatic Cell and Molecular Genetics

Abstract

A somatic cell hybrid line, called M11-X, was developed in order to study the expression and regulation of the human β-like globin genes in a mouse erythroid environment. M11-X cells were obtained by fusing the human fibroblast cell line GM3552 (which contains the translocation chromosome t(11;X) that carries the human β-like globin genes) with hypoxanthine phosphoribosyltransferase (HPRT)-negative tetraploid (2S) mouse erythroleukemia (MEL) cells. After induction with 5 mM hexamethylene bisacetamide (HMBA), these cells contain approximately 300–600 copies per cell of correctly initiated, processed, and terminated human β-globin mRNA; however, neither human ε-nor γ -globin mRNAs were detected. Carboxymethylcellulose chromatography followed by SDS-polyacrylamide gel electrophoresis and Western blotting revealed that normal human β-globin protein was also present. These results suggest that the human β-globin gene, when present in mouse erythroid cells, can be transcribed and its mRNA translated into normal products, but at a much lower level than the mouse β-globin genes. Analysis of the frequency of cytosine methylation near the human γ-globin genes indicated that these genes are heavily methylated in M11-X cells. The inability to express the human γ-globin genes of these cells might be accounted for, at least in part, by DNA methylation.

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Author notes

  1. Linda Sanders-Haigh

    Present address: Department of Biology, Massachusetts Institute of Technology, 02139, Cambridge, Massachusetts

Authors and Affiliations

  1. Laboratory of Molecular Hematology, National Heart, Lung, and Blood Institute, National Institutes of Health, 20205, Bethesda, Maryland

    Yawen L. Chiang, Linda Sanders-Haigh & W. French Anderson

  2. Clinical Hematology Branch, National Heart, Lung, and Blood Institute, National Institutes of Health, 20205, Bethesda, Maryland

    Timothy J. Ley

Authors
  1. Yawen L. Chiang
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  2. Timothy J. Ley
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  3. Linda Sanders-Haigh
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  4. W. French Anderson
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Chiang, Y.L., Ley, T.J., Sanders-Haigh, L. et al. Human globin gene expression in hybrid 2S MEL × human fibroblast cells. Somat Cell Mol Genet 10, 399–407 (1984). https://doi.org/10.1007/BF01535635

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  • DOI: https://doi.org/10.1007/BF01535635

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