Introduction to the Workshop
URLs Provided by Attendees

Abstracts

Mapping

Informatics

Sequencing

Instrumentation

Ethical, Legal, and Social Issues

Infrastructure

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.

An Integrated Map of Human Chromosome 16

N.A. Doggett[1], D.F. Callen[2], M.R. Altherr[1], L.A. Duesing[1], J.G.Tesmer[1], L.J. Meincke[1], D.C. Bruce[1], A.A. Ford[1], D.C. Torney[1], R.D. Sutherland[1], M.G. Lowenstein[1], M.O. Mundt[1], W.J. Bruno[1], E.H. Knill[1], R.I. Richards[2], G.R. Sutherland[2], L.L. Deaven[1], and R.K. Moyzis[1]
[1]Life Sciences Division and Center for Human Genome Studies, Los Alamos National Laboratory, Los Alamos, NM, [2]Cytogenetics and Molecular Genetics, Woman's & Children's Hospital, Adelaide, SA, Australia

We have constructed an integrated physical/genetic/cytogenetic map of human chromosome 16. The framework for constructing this map is a high resolution cytogenetic breakpoint map derived from 78 mouse/human somatic cell hybrids and 4 fragile sites which divide chromosome 16 into 90 intervals of average size 1 Mb. The physical map consists of both a low resolution YAC contig map and a high resolution cosmid contig map. The low resolution YAC contig map is comprised of 515 CEPH MegaYACs, and 220 flow sorted 16-specific YACs that are localized to and ordered within the breakpoint intervals with 320 STSs. This YAC map provides nearly complete coverage of the euchromatic arms of the chromosome.

A high resolution "sequence ready" cosmid contig map consisting of 4000 fingerprinted cosmids assembled into contigs covering 60% of the chromosome is anchored to the YAC and cytogenetic breakpoint maps via STSs developed from cosmid contigs and by hybridizations between YACs and cosmids. To date, 300 of these contigs containing greater than 1600 cosmids have been merged with the integrated physical/genetic/cytogenetic map and thus regionally localized along the chromosome.

A highly informative microsatellite-based genetic map (developed at the Adelaide Children's Hospital) consisting of 78 PCR typeable markers and having a 2.6 cM median (3.2 average) distance between markers is tightly integrated with the physical map because most of these GT/CA markers were developed from localized cosmids or screened against the YAC map, and by regional localization of all markers to the cytogenetic breakpoint map.

To integrate a transcription map of chromosome 16 with the existing physical/genetic/cytogenetic map, we have subjected 14,000 chromosome 16 cosmids to exon amplification, isolated 5000 exon clones and sequenced 500 of these clones. Over 500 genes, anonymous DNA markers and microsatellite repeats have also been incorporated as part of an ongoing effort to integrate all available GDB loci with the 16 map. This integrated map is facilitating the cloning of various disease genes and fragile sites on chromosome 16.

Supported by the US DOE (W-7405-ENG-36).