Introduction to the Workshop
URLs Provided by Attendees
- Ethical, Legal, and Social Issues
The electronic form of this document may be cited in the following style:
Human Genome Program, U.S. Department of Energy, DOE Human Genome Program Contractor-Grantee Workshop IV, 1994.
Abstracts scanned from text submitted for November 1994 DOE Human Genome Program Contractor-Grantee Workshop. Inaccuracies have not been corrected.
Development of a Multi Level Resolution Map of the p arm of Chromosome 19 by Integration of FISH, YAC, Cosmid Contig, and Restriction Map Data
Susan A. Allen, Anca M. Georgescu, Maria T. de Jesus, Susan M. G. Hoffman, Gregory G. Lennon, Anthony V. Carrano and Anne S. Olsen
Human Genome Center, L-452, Biology and Biotechnology Research Program, Lawrence Livermore National Laboratory, Livermore, California 94550.
A comprehensive map of the p arm of chromosome 19 is being constructed integrating data from various methods. Each form of data contributes to the integrated map at different levels of resolution. An initial framework map was developed by fluorescence in situ hybridization (FISH) of 80 cosmid markers for which order and relative distance was determined along the 20 Mb span of 19p (see poster by Brandriff et al). Most of the ordered cosmid markers are members of cosmid contigs assembled by the fingerprinting procedure that constitutes the foundation of the chromosome 19 map. Cosmid contigs in close proximity were often merged by directed walking in cosmid, BAC, PAC or P1 libraries. In addition, cosmid walking often incorporated previously unmapped contigs into the map.
Remaining gaps in the cosmid contig map are initially being spanned by YACs isolated by STS screening and Alu-PCR hybridization. At present, a total of 106 STSs, including 26 polymorphic markers, have been developed from ordered cosmids or derived from outside sources. These STSs have been subsequently integrated into the map (see poster by Tsujimoto et al). Over 90 YACs have been added to the ordered map, thereby reducing the 80 ordered cosmid contigs to less than 36 YAC-cosmid islands. These islands span an estimated 15 Mb or about 75% of the p arm. Alu-PCR products from YACs have been hybridized to cosmid colony filters to incorporate additional cosmid contigs into the map and to close gaps between YACs.
At the highest level of resolution, EcoRI restriction maps have been constructed for cosmid contigs spanning an accumulated 5.6 Mb (see poster by Burgin et al). This serves to verify the established contigs and determine the length of the DNA spanned by overlapping clones. This procedure will eventually localize individual genes and genetic markers with an average resolution of 10 Kb.
This work was performed under the auspices of the U.S. Department of Energy by Lawrence Livermore National Laboratory under contract no. W-7405-ENG-48.