Transcriptional mapping by direct cDNA selection: From Xq28 to the entire X chromosome
- Deutsches Krebsforschungszentrum, Heidelberg (Germany)
The human X chromosomes has been studied by genetic and molecular means for a long time. The terminal part of the long arm of the chromosome has been screened for transcripts in every detail by cDNA selection. We were able to re-isolate all known genes from this region out of cDNA sublibraries generated by using genomic clones as templates for the selection technology. Overall we used about 600 cosmids, 2 P1 clones and 7 YACs to generate more than 20,000 cDNA clones, which are analyzed by hybridization and exclusion sequencing. The complexity of our cDNA source (from three human tissues) seems to be very high, as so far we have not identified any clones which are identical. Due to mapping randomly picked clones, we estimate that more than 80% of the cDNA clones do map back to the chromosomal region used for cDNA selection. We are now in the process of constructing cDNA sublibraries from the entire X chromosome and other chromosomes, using modified protocols. We currently use cDNA from six different human tissues. The genomic templates are either flow sorted chromosomes or pools of genomic clones. Mapping of the selected cDNA clones is performed on characterized YAC clones of the X chromosome, hybrid cell line panels or by FISH. The rapidly growing transcriptional maps provide candidate genes for genetic diseases which are mapped along the chromosome. The analysis will be complemented by exon trapping (internal and 3{prime}) of specific regions. In combination with other mapping approaches on the X chromosome it will be possible to construct an integrated physical, genetic and transcript map.
- OSTI ID:
- 134426
- Report Number(s):
- CONF-941009-; ISSN 0002-9297; TRN: 95:005313-1160
- Journal Information:
- American Journal of Human Genetics, Vol. 55, Issue Suppl.3; Conference: 44. annual meeting of the American Society of Human Genetics, Montreal (Canada), 18-22 Oct 1994; Other Information: PBD: Sep 1994
- Country of Publication:
- United States
- Language:
- English
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