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.

Using Expressed Sequences as Nucleation Points for Genomic Sequencing

Michael R. Altherr, Amanda Ford, Cleo Naranjo, Judy Buckingham, Chris Munk and Robert K. Moyzis.
Life Sciences Division and Center for Human Genome Studies, Los Alamos National Laboratory, Los Alamos, NM 87545.

Individual chromosomes provide the skeletal framework on which genomic data is organized. As the physical map and an ordered assemblage of molecular clones for chromosome 16 nears completion, we have embarked on the construction of an 'expressed sequence map' of this chromosome. Assuming that there are 100,000 human genes, approximately 3,000 should be encoded by chromosome 16. To this end, we have chosen the strategy of exon amplification to identify expressed sequences on chromosome 16. Our strategy employs 96-well plate pools of DNA from the flow sorted and arrayed chromosome 16 cosmid library as the substrate for exon trapping. At present we have generated 1800 exon clones from 75 of the 150 plates in the chromosome 16 library. We have sequenced over 500 of these clones and determined that approximately 60% appear to be independent exons. These sequences are being mapped to specific locations on chromosome 16 using a panel of somatic cell hybrids, YACs and cosmids that have previously been integrated into our high resolution physical map. In addition, these sequences are being subjected to database analysis to determine whether they represent previously characterized genes or contained conserved motifs that might provide some insight as to their biological function. The exonic sequences obtained in this way can be used as nodes from which to initiate genomic sequencing efforts. We have used a 230 kbp cosmid contig from the 7q telomere to model this integrated exon amplification/genomic sequencing approach.