of Work: The substantial advancements in TAR cloning in the Chromosome Stability Group provided for development of a separate, independent Gene Isolation Unit that devotes much of its efforts to refining TAR cloning approaches for mammalian tissues. We initiated a collaboration with Ron Walter, Southwest Texas University, to develop yeast artificial libraries (YAC) from the fish Xiphophorous, since these fish are excellent whole animal genetic models for DNA repair and tumor suppressor control. We have also started the isolation of regions surrounding specific genes where known tumor suppressor genes have been mapped (Xmrk and Cdkn2 regions). TAR cloning of cDNAs is being refined to examine expressed genes from various cancer tissues using our yeast functional assays. In other studies we have shown that TAR cloning of p53 cDNA from human cell lines and cancer tissue is an accurate process and that yeast-based systems can be used to characterize mutant p53s. Based on the results of the TAR cloning of the human p53, we have initiated a project to clone and test the p53 from Xiphophorous, and then to test p53's from different induced tumors in our yeast based system.

Agency
National Institute of Health (NIH)
Institute
National Institute of Environmental Health Sciences (NIEHS)
Type
Intramural Research (Z01)
Project #
1Z01ES065072-10
Application #
6432378
Study Section
(LMG)
Project Start
Project End
Budget Start
Budget End
Support Year
10
Fiscal Year
2000
Total Cost
Indirect Cost
Name
U.S. National Inst of Environ Hlth Scis
Department
Type
DUNS #
City
State
Country
United States
Zip Code
Storici, Francesca; Bebenek, Katarzyna; Kunkel, Thomas A et al. (2007) RNA-templated DNA repair. Nature 447:338-41
Storici, Francesca; Resnick, Michael A (2006) The delitto perfetto approach to in vivo site-directed mutagenesis and chromosome rearrangements with synthetic oligonucleotides in yeast. Methods Enzymol 409:329-45
Storici, Francesca; Snipe, Joyce R; Chan, Godwin K et al. (2006) Conservative repair of a chromosomal double-strand break by single-strand DNA through two steps of annealing. Mol Cell Biol 26:7645-57
Storici, Francesca; Resnick, Michael A (2003) Delitto perfetto targeted mutagenesis in yeast with oligonucleotides. Genet Eng (N Y) 25:189-207
Storici, Francesca; Durham, Christopher L; Gordenin, Dmitry A et al. (2003) Chromosomal site-specific double-strand breaks are efficiently targeted for repair by oligonucleotides in yeast. Proc Natl Acad Sci U S A 100:14994-9
Storici, F; Lewis, L K; Resnick, M A (2001) In vivo site-directed mutagenesis using oligonucleotides. Nat Biotechnol 19:773-6
Humble, M C; Kouprina, N; Noskov, V N et al. (2000) Radial transformation-associated recombination cloning from the mouse genome: isolation of Tg.AC transgene with flanking DNAs. Genomics 70:292-9