Construction of a YAC contig in the region of three retinal degeneration genes in 11q13
- Washington Univ. School of Medicine, St. Louis, MO (United States)
We have previously isolated a large number of bacteriophage clones specific for human chromosome 11. Seventy-two clones map to distal 11q13, a region haboring a number of human disease genes. Among them are genes for Best macular degeneration (BMD), Usher`s syndrome type 1B, autosomal dominant neovascular inflammatory vitreoretinopathy (ADNIV) and a form of Bardet-Biedl syndrome; all diseases involve retinal degenerations. From the genetic mapping it is clear the BMD and Bardet-Biedl syndrome are located in the proximal portion of the band, while Usher`s syndrome 1B and ADNIV are in the distal portion of 11q13. As the first step towards the cloning of the latter two disease genes, we set out to isolate this region of the chromosome as yeast artificial chromosomes (YACs). The 72 clones, as well as two transcribed genes, were assigned into 4 segregation groups with radiation-reduced somatic cell hybrids. We have shown previously that this high probe density can result in the development of large YAC contigs. We chose to use two complementing strategies to screen three different YAC libraries. The two strategies are: (1) the generation of Alu-primed products from the bacteriophage clones that are then used as probes against filters from individual YAC library pools; (2) the sequencing of the bacteriophage clones, STS development and screening YAC libraries by the method of Green and Olson (1990). The former has the advantage of speed, while the latter of specificity. We have identified a total of 140 YACs from 50 screens. Approximately 50% of the markers are already in small contigs. The largest contig has 10 markers and is {approximately}1 mbp. This approach results in fairly rapid development of YAC contigs without additional application of labor-intensive, locus-specific manipulation. We expect to complete the contig within a year.
- OSTI ID:
- 134408
- Report Number(s):
- CONF-941009--
- Journal Information:
- American Journal of Human Genetics, Journal Name: American Journal of Human Genetics Journal Issue: Suppl.3 Vol. 55; ISSN AJHGAG; ISSN 0002-9297
- Country of Publication:
- United States
- Language:
- English
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