Michael R. Altherr, Darrell Ricke. Amanda Ford. Cleo Naranjo. Jason Collins. Michael Lowenstein. Norman Doggett. Larry Deaven and Robert K. Moyzis.
Life Sciences Division and Center for Human Genome Studies, Los Alamos National Laboratory, Los Alamos, NM.
Individual chromosomes provide the skeletal framework on which genetic data are organized. As the physical map and an ordered assemblage of molecular clones for chromosome 16 neared completion (Doggett, et al., Nature 377 Suppl.:335), we 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. The strategy employed 96-well plate pools of DNA from the flow sorted and arrayed chromosome 16 cosmid library as the substrate for exon trapping. We have generated and archived more than 3,024 exon clones from 126 plates in the chromosome 16 library. We have sequenced over 2000 of these clones and determined that approximately 50% are distinct. These sequences were analyzed using available databases and are being mapped to specific locations on chromosome 16 using a grided array of previously fingerprinted cosmids that are integrated into a high resolution physical map. We anticipate that this effort will provide a 1000 member exon map of chromosome 16 as the first step toward a chromosome 16 gene map. This effort is complemented by the sample sequencing (SASE) approach to gene identification on chromosome 16 (see Han et al. and Ricke et al. this meeting).
This work was supported by the Department of Energy under contract W7405-EMG-36.
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