Monochromosomal hybrids for the analysis of the human genome. Final report
The objective of this research project was to produce a panel of mouse/human hybrid cell lines each harboring a single different human chromosome. The human chromosome present in rodent cells was marked with a dominant selectable marker and maintained by selection. In studies, an amphotropic retroviral vector pZIPgpt that carries a dominant selectable marker gpt, was used to tag chromosomes in normal diploid cells. Human DNA flanking the integrated vector was obtained by PCR amplification and used to probe Southern blots of DNAs of a hybrid cell panel to identify the chromosome of its origin. This allowed them to generate clonal human cell lines, each carrying the gpt integrated into a different chromosome. Human chromosomes bearing the selectable marker are then transferred to mouse cells by the microcell fusion method. The authors have produced a monochromosomal hybrid panel comprised of human chromosomes 1 through 21 and X. A monochromosomal hybrid for chromosome 15 without a selectable marker has also been obtained. Chromosomes missing in the panel include 22 and Y. All of the hybrid cell lines have been characterized by chtogenetic and molecular methods to ascertain the identity and structural integrity of the human chromosome. A panel of monochromosomal hybrid cell lines has been used for complementation analysis, cloning of chromosome specific DNA fragments, physical mapping of human chromosomes and fractionation of the human genome.
- Research Organization:
- Temple Univ., Philadelphia, PA (United States). Fels Inst. for Cancer Research and Molecular Biology
- Sponsoring Organization:
- USDOE Office of Energy Research, Washington, DC (United States)
- DOE Contract Number:
- FG02-94ER61750
- OSTI ID:
- 453481
- Report Number(s):
- DOE/ER/61750--T1; ON: DE97003737
- Country of Publication:
- United States
- Language:
- English
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