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Research Article

Isolation of microcell hybrid clones containing retroviral vector insertions into specific human chromosomes.

T G Lugo, B Handelin, A M Killary, D E Housman, R E Fournier
T G Lugo
Department of Microbiology, University of Southern California School of Medicine, Los Angeles 90033.
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B Handelin
Department of Microbiology, University of Southern California School of Medicine, Los Angeles 90033.
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A M Killary
Department of Microbiology, University of Southern California School of Medicine, Los Angeles 90033.
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D E Housman
Department of Microbiology, University of Southern California School of Medicine, Los Angeles 90033.
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R E Fournier
Department of Microbiology, University of Southern California School of Medicine, Los Angeles 90033.
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DOI: 10.1128/MCB.7.8.2814
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ABSTRACT

We sought an efficient means to introduce specific human chromosomes into stable interspecific hybrid cells for applications in gene mapping and studies of gene regulation. A defective amphotropic retrovirus was used to insert the gene conferring G418 resistance (neo), a dominant selectable marker, into the chromosomes of diploid human fibroblasts, and the marked chromosomes were transferred to mouse recipient cells by microcell fusion. We recovered five microcell hybrid clones containing one or two intact human chromosomes which were identified by karyotype and marker analysis. Integration of the neo gene into a specific human chromosome in four hybrid clones was confirmed by segregation analysis or by in situ hybridization. We recovered four different human chromosomes into which the G418 resistance gene had integrated: human chromosomes 11, 14, 20, and 21. The high efficiency of retroviral vector transformation makes it possible to insert selectable markers into any mammalian chromosomes of interest.

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Isolation of microcell hybrid clones containing retroviral vector insertions into specific human chromosomes.
T G Lugo, B Handelin, A M Killary, D E Housman, R E Fournier
Molecular and Cellular Biology Aug 1987, 7 (8) 2814-2820; DOI: 10.1128/MCB.7.8.2814

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Isolation of microcell hybrid clones containing retroviral vector insertions into specific human chromosomes.
T G Lugo, B Handelin, A M Killary, D E Housman, R E Fournier
Molecular and Cellular Biology Aug 1987, 7 (8) 2814-2820; DOI: 10.1128/MCB.7.8.2814
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