A method for generating hybrids containing nonselected fragments of human chromosomes

Journal Article · Mon May 01 04:00:00 EDT 1989 · Genomics; (USA)

DOI:https://doi.org/10.1016/0888-7543(89)90274-7· OSTI ID:5401506

Benham, F; Hart, K; Crolla, J; Bobrow, M; Francavilla, M; Goodfellow, P N ^[1]

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1. United School of Guy's Hospital, London (England)

We have used an irradiation and fusion technique to generate somatic cell hybrids that contain human chromosomal fragments. As a model system, a human-hamster hybrid containing a single human X chromosome was gamma-irradiated and fused with a rodent line. Hybrids were obtained without imposing direct selection for human material. Analysis of 29 clones by in situ hybridization and Southern blotting revealed that human fragments were incorporated into the hybrid cell genomes in most lines. Like chromosome-mediated gene transfer (CMGT)-generated hybrids, these hybrids contained multiple human fragments and retained alphoid centromeric sequences with a high frequency. However, unlike the CMGT, human fragments (apart from alphoid sequences) of less than 10(7) bp showed no evidence for rearrangements. This technique provides a method for constructing hybrids that contain a limited number of small human fragments derived exclusively from any chromosome of choice without the need to impose selection. Such hybrids provide a valuable resource for high-resolution mapping over short distances and for the isolation of disease and other loci mapped genetically.

• OSTI ID: 5401506
• Journal Information: Genomics; (USA), Journal Name: Genomics; (USA) Vol. 4:4; ISSN GNMCE; ISSN 0888-7543
• Country of Publication: United States
• Language: English

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