High density transcriptional mapping of chromosome 21 by hybridization selection
- Eleanor Roosevelt Institute, Denver, CO (United States); and others
A transcriptional map of human chromosome 21 is important for the study of Down syndrome, development processes and genome organization. To construct a high density transcriptional map, the technique of cDNA hybrid selection is being applied to a minimal tiling path of YAC clones that span 21q. The cDNA used for selection represents a complex pool of sequences obtained from a variety of fetal and adult tissues and cell lines. Approximately 70-80 YAC clones are sufficient to span 21q; each is individually processed through the selection procedure to obtain a YAC-specific {open_quotes}selected cDNA library{close_quotes}. Survey analysis of each library includes determination of levels of ribosomal contamination, verification of enrichment of control genes, identification of a preliminary number of novel unique sequences, and verification that novel sequences map to the correct YAC and chromosomal regions. This analysis has been completed for 19 YACs that together comprise approximately 10 Mb of non-overlapping DNA, 25% of the long arm. Ribosomal cDNA contamination is low (<10%) and all known genes of appropriate tissue specificity of expression have been recovered, as well as new genes from each YAC. Libraries of expression have been recovered, as well as new genes from each YAC. Libraries from 8 of these YACs are now being subjected to exhaustive analysis to identify all novel genes contained within them and to obtain complete cDNAs and expression analysis for each. Not all regions of the chromosome, however, are equally amenable to these analyses. Selected cDNA libraries from the centromeric YACs are yielding apparently novel genes, but confirmation of map position is problematic. Also of interest is a region of several megabases within the Giemsa dark band, 21q21. Selected cDNA libraries from these YACs so far have yielded no novel genes and support the idea of a genuinely very gene-poor region.
- OSTI ID:
- 134483
- 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
Similar Records
YAC contig and cell hybrid mapping of six expressed sequences encoded by human chromosome 21
Discovering, mapping and characterization of 39 new genes in human MHC class I region by cDNA selection