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Transfer of the human HPRT and GART genes from yeast to mammalian cells by microinjection of YAC DNA

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Somatic Cell and Molecular Genetics

Abstract

DNA of two yeast artificial chromosomes (YACs) containing selectable human genes was transferred by microinjection to rodent cells in tissue culture. The human hypoxanthine phosphoribosyltransferase (HPRT) gene, spanning 45 kb, is contained on the 660-kb YAC yHPRT as described elsewhere. The human phosphoribosylglycinamide formyltransferase (GART) gene, spanning approximately 40 kb, is contained on the 590-kb YAC yGART2 as described previously. YAC DNA was isolated from pulsed-field gels and microinjected into mammalian cells in which the human HPRT and GART genes can be selected. The cell lines that were selected contain the entire human genes. Some of the cell lines contain multiple copies of the genes integrated at the same chromosomal position. The YAC yGART2 could not be purified away from natural yeast chromosomes of similar size, and the cell lines into which the human GART gene was introduced contain variable amounts of yeast DNA in addition to the human DNA.

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Authors and Affiliations

  1. Department of Genetics, Washington University School of Medicine, 63110, St. Louis, Missouri

    Andreas Gnirke & Clare Huxley

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  1. Andreas Gnirke
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  2. Clare Huxley
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Gnirke, A., Huxley, C. Transfer of the human HPRT and GART genes from yeast to mammalian cells by microinjection of YAC DNA. Somat Cell Mol Genet 17, 573–580 (1991). https://doi.org/10.1007/BF01233622

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  • DOI: https://doi.org/10.1007/BF01233622

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