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Genetics, Vol 141, 147-157, Copyright © 1995
INVESTIGATIONS |
Integrated Maps of the Chromosomes in Dictyostelium discoideum
W. F. Loomis, D. Welker, J. Hughes, D. Maghakian and A. Kuspa
Center for Molecular Genetics, Department of Biology, University of California at San Diego, La Jolla, California 92093
Detailed maps of the six chromosomes that carry the genes of Dictyostelium discoideum were constructed by correlating physically mapped regions with parasexually determined linkage groups. Chromosomally assigned regions were ordered and positioned by the pattern of altered fragment sizes seen in a set of restriction enzyme mediated integration-restriction fragment length polymorphism (REMI-RFLP) strains each harboring an inserted plasmid that carries sites recognized by NotI, SstII, SmaI, BglI and ApaI. These restriction enzymes were used to digest high molecular weight DNA prepared from more than 100 REMI-RFLP strains and the resulting fragments were separated and sized by pulsed-field gels. More than 150 gene probes were hybridized to blots of these gels and used to map the insertion sites relative to flanking restriction sites. In this way, we have been able to restriction map the 35 mb genome as well as determine the map position of more than 150 genes to with ~40 kb resolution. These maps provide a framework for subsequent refinement.
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