Ellen Peterson. Donna L. Robinson, Leslie Chasteen, Robert Sutherland, Linda S. Thompson, Meryl Gersh, Joan Overhauser, Larry L. Deaven, Robert K. Moyzis, and Deborah L. Grady
Center for Human Genome Studies and Life Sciences Division, Los Alamos National Laboratory, Los Alamos, NM, and Thomas Jefferson University, Philadelphia, PA.
A total of 304 new STSs have been generated from flow sorted human chromosome 5 DNA. These STSs have been regionally ordered using breakpoint analysis to one of 51 bins on 5p and to one of 16 bins on 5q. The current density of markers (1/640 kb), in addition to the numerous PCR based genetic markers generated by other groups, is sufficient to provide nucleation points for YAC contig assembly in all regions of chromosome 5. Complete Mega-YAC contigs have been generated for the critical region (5p15.2) [Genomics 24(1):63-8, 1994] and the cat-cry region (5p15.3) identified with the cri-du-chat syndrome on the short arm of chromosome 5. This work has been extended to include YAC coverage of the entire short arm of this chromosome. The short arm constitutes approximately 50 Mb of the total 194 Mb of DNA on chromosome 5. This map has been generated by STS content mapping of YACs from the published Genethon tiling data set and by direct screening of the entire Genethon library. YACs with a high probability of containing these new STSs were collected. This probability was established by binning Genethon STSs on the same hybrid panel used on the chromosome 5 STS data. YACs localized to a specific bin by Genethon STSs were likely to contain chromosome 5 specific STSs in that same bin. By systematically testing our STSs by PCR against probable YACs and screening the library for novel YACs, a YAC contig was constructed. Currently the contig, covering >90% of 5p, consists of 531 mega-YACs and 161 integrated STSs. The completion of this map will serve as a resource for the identification of other genes on the short arm of human chromosome 5, and form the framework to construct a high resolution E. coli based map for eventual DNA sequencing.
* This work was supported by USDOE under contract W-7405-ENG-36.