Genetic and physical mapping of human and animal genomes has been greatly facilitated by using chromosome-specific DNA libraries. Although DNA libraries for most of the human chromosomes have been constructed, little has been documented for farm animals. The present study was undertaken to establish approaches for mapping single chromosomes. For this purpose, three experiments were carried out: microcell-mediated chromosome transfer; chromosome microdissection and cloning; and physical mapping of chicken chromosome 1 with chromosome 1-specific cosmid clones. In the first experiment, microcells prepared from pSV2-neo plasmid transfected chicken cells were fused to human HeLa cells. Three hybrid cell lines, each carrying a single chicken chromosome or a fragment of a chicken chromosome were established. In the second experiment, the chicken chromosome 1 was isolated by microdissection. The chromosomal DNA was amplified by PCR and DNA in microgram quantities was obtained. A chicken chromosome 1-specific DNA library was constructed from the PCR amplified DNA through conventional cloning. In the third experiment, a chicken genomic DNA cosmid library was constructed. Using chicken chromosome 1 specific DNA as a probe, aliquots of the cosmid library were screened and 6 cosmids located in different regions on chicken chromosome 1 were isolated and physically mapped to the chromosome by in situ hybridization. This study has demonstrated the applicability of the microcell-mediated chromosome transfer and chromosome microdissection for isolation of chromosome-specific DNA. The chicken chromosome 1-specific DNA library and chromosome assignment of chromosome specific cosmid clones will facilitate the development of a linkage map of the chicken genome.
| Identifer | oai:union.ndltd.org:UMASS/oai:scholarworks.umass.edu:dissertations-8977 |
| Date | 01 January 1994 |
| Creators | Li, Yukui |
| Publisher | ScholarWorks@UMass Amherst |
| Source Sets | University of Massachusetts, Amherst |
| Language | English |
| Detected Language | English |
| Type | text |
| Source | Doctoral Dissertations Available from Proquest |
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