During the next granting period, our goals will focus on the utilization of the cloned genomic DNA sequences in YAC contigs from the short arm of chromosome 11 to develop a detailed map of expressed genes for this chromosome arm. In order to carry out this objective our specific goals will be as follows: 1. We will complete YAC contig generation for the short arm of chromosome 11. To achieve this goal we will continue to implement the alu pcr based filter hybridization strategy we have devised to achieve a high level of efficiency and economy in YAC contig building. 2. We will characterize the genomic DNA isolated by the YAC cloning to verify the accuracy with which genomic DNA sequences from human DNA are represented in the YACs 3. We will utilize DNA from 11p YAC contigs to identify sequences of expressed genes in the genomic DNA of this chromosome arm. To achieve this objective efficiently we will continue to implement the exon amplification methodology we developed during the previous granting period. 4. We will isolate cDNAs for each expressed gene we identify. To facilitate this goal we will implement pcr based cDNA library screening strategies to maximize efficiency of library screening. 5. We will attempt to develop and implement an efficient strategy for obtaining full length cDNA coverage for each gene. 6. We will carry out sequence analysis of cDNAs and corresponding segments of genomic DNA. 7. We will develop YAC contigs in the mouse for the genomic regions homologous to the 11p regions on which we have focused. We will utilize these YAC contigs in order to develop a comparative map of expressed genes for the syntenic chromosome segments between the two species.
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