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PMID: 2823107 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

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

Molecular and cellular biology ·Vol. 7 ·No. 8 ·1987-08-00 ·Pages 2814-20

Lugo TG, Handelin B, Killary AM, Housman DE, Fournier RE

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.

MeSH Terms
Animals Cell Transformation, Neoplastic Cells, Cultured Chromosomes, Human Clone Cells DNA Transposable Elements Humans Karyotyping Metaphase Mice Retroviridae/genetics
Chemicals
DNA Transposable Elements
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Lugo T G
Department of Microbiology, University of Southern California School of Medicine, Los Angeles 90033.
Handelin B
Killary A M
Housman D E
Fournier R E
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.7.8.2814-2820.1987
Published
1987-08-00
Pages
2814-20
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC367898
Subset
IM
Grants
NIGMS NIH HHS · GM 26449 · United States
External Links
PubMed source DOI: 10.1128/mcb.7.8.2814-2820.1987 PMC Full Text View journal details
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