Abstract

This renewal application is focused upon two issues that are central to finding genes in the human genome. The first of these relates to the development of technology refinements to improve the selection and extension of cDNAa. Full length cDNAs are valuable tools in the positional cloning of disease genes, for annotating genomic DNA sequences and for functional genomics. However, no quick routes currently exist for moving from a cDNA fragment to a full length cDNA. We propose developing enhancements to increase the length of direct selected cDNAs and to enable the rapid conversion of cDNAs to full length. Direct cDNAs selection has been successfully applied to entire human chromosomes. We now propose applying these techniques to derive normalized cDNAs using the whole human genome. These types of cDNA libraries would complement and enhance existing normalized cDNA resources. The second issue that this application addresses is the development and mapping of new Expressed Sequence Tags (ESTs). The catalogue of human genes is far from complete, and gene prediction algorithms based upon genomic DNA sequences are not fully reliable. We propose to build a detailed map of 2,000 genes on human chromosomes 5 which is biased towards sequences that are not in the EST database. This will be facilitated by techniques we have developed that enrich for cDNAs which other methods miss. In our final aim we propose a limited scale up of this approach to derive new chromosome-specific cDNA libraries and novel EST maps for human chromosomes 3 and 8.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Human Genome Research Institute (NHGRI)
Type
Research Project (R01)
Project #
7R01HG000368-11
Application #
2889646
Study Section
Special Emphasis Panel (ZRG2-BIOL-1 (01))
Program Officer
Feingold, Elise A
Project Start
1988-09-30
Project End
2001-06-30
Budget Start
1999-07-01
Budget End
2001-06-30
Support Year
11
Fiscal Year
1999
Total Cost
Indirect Cost

  Institution

Name
Washington University
Department
Genetics
Type
Schools of Medicine
DUNS #
062761671
City
Saint Louis
State
MO
Country
United States
Zip Code
63130

  Publications

Boyadjiev, S A; Chowdry, A B; Shapiro, R E et al. (2002) Physical map of the chromosome 6q22 region containing the oculodentodigital dysplasia locus: analysis of thirteen candidate genes and identification of novel ESTs and DNA polymorphisms. Cytogenet Genome Res 98:29-37
Rojas, K; Serrano de la Pena, L; Gallardo, T et al. (1999) Physical map and characterization of transcripts in the candidate interval for familial chondrocalcinosis at chromosome 5p15.1. Genomics 62:177-83
Hastbacka, J; Kerrebrock, A; Mokkala, K et al. (1999) Identification of the Finnish founder mutation for diastrophic dysplasia (DTD). Eur J Hum Genet 7:664-70
Bardien-Kruger, S; Greenberg, J; Tubb, B et al. (1999) Refinement of the RP17 locus for autosomal dominant retinitis pigmentosa, construction of a YAC contig and investigation of the candidate gene retinal fascin. Eur J Hum Genet 7:332-8
Simmons, A D; Overhauser, J; Lovett, M (1997) Isolation of cDNAs from the Cri-du-chat critical region by direct screening of a chromosome 5-specific cDNA library. Genome Res 7:118-27
Touchman, J W; Bouffard, G G; Weintraub, L A et al. (1997) 2006 expressed-sequence tags derived from human chromosome 7-enriched cDNA libraries. Genome Res 7:281-92
Wise, C A; Clines, G A; Massa, H et al. (1997) Identification and localization of the gene for EXTL, a third member of the multiple exostoses gene family. Genome Res 7:10-6
Clines, G A; Ashley, J A; Shah, S et al. (1997) The structure of the human multiple exostoses 2 gene and characterization of homologs in mouse and Caenorhabditis elegans. Genome Res 7:359-67
Stickens, D; Clines, G; Burbee, D et al. (1996) The EXT2 multiple exostoses gene defines a family of putative tumour suppressor genes. Nat Genet 14:25-32
Guimera, J; Casas, C; Pucharcos, C et al. (1996) A human homologue of Drosophila minibrain (MNB) is expressed in the neuronal regions affected in Down syndrome and maps to the critical region. Hum Mol Genet 5:1305-10

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