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Related Experiment Videos

Rat growth hormone expression in cell hybrids.

J S Strobl, R Padmanabhan, B H Howard

    DNA (Mary Ann Liebert, Inc.)
    |January 1, 1984
    PubMed
    Summary

    Rat growth hormone (rGH) gene expression is silenced in hybrid cells due to mouse cell factors. Introducing the rGH gene into these hybrids restores gene expression, suggesting repressors don't target the introduced gene.

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    Area of Science:

    • Molecular Biology
    • Genetics
    • Cell Biology

    Background:

    • Rat growth hormone (rGH) gene expression is extinguished in somatic cell hybrids formed from GH3 (rat pituitary adenoma) and L (mouse fibroblast) cells.
    • This extinction is hypothesized to result from transacting factors present in the L cells.

    Purpose of the Study:

    • To investigate the mechanisms of rGH gene silencing in hybrid cells.
    • To determine if introduced rGH genes can overcome silencing and restore gene expression.

    Main Methods:

    • Cloned rGH gene was introduced into hybrid cells via microinjection and calcium phosphate transfection.
    • Gene expression was assessed using immunofluorescence for GH peptides and Northern hybridization for rGH transcripts.

    Main Results:

    • GH peptides and normal-sized rGH mRNA transcripts were detected in transfected hybrid cells.
    • Stable expression of rGH was achieved with as few as one or two additional gene copies.
    • Expression occurred even in hybrid cells lacking the endogenous rGH gene.

    Conclusions:

    • Transacting repressors in hybrid cells do not appear to recognize the introduced rGH genes.
    • Hybrid cells possess elements that permit the production of functional, normal-sized rGH mRNA, unlike L cells.

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