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Content archived on 2024-04-30

Construction of an integrated transcriptional map of the human X chromosome (2)


- - To construct a cosmid/PAC/BAC map of the human X chromosome with reference clones accessible to all participants and based on highly annotated YAC maps.
- - To isolate and map the majority of X chromosome transcripts to the cosmide pockets and when established, to the cosmid contigs.
- - To perform large scale sequencing of selected regions of the X chromosome, principally Xq28 and Xp22.

With the availability of near complete coverage of the human X chromosome in YAC contigs, the participants aim at a coordinated effort to construct a map of bacterial vectors, as a substrate to the isolation of most of the transcripts of the chromosome. For both objectives, two different and complementary strategies will be used. First a global and systematic approach will be used by the coordinator. A minimal tiling path of YAC DNA will be isolated by long-range Alu-PCR and screened against a variety of reference cosmid, PAC and BAC libraries, thus constructing a pocket map of the chromosome. A cDNA library enriched for X chromosome transcripts will be constructed by laboratory in Heidelberg and made available to all participants. It will be systematically screened with cosmids from the pocket map to position transcripts. Secondly, a more regional approach will be used by key participants on the X chromosome, who have accumulated large amounts of material and expertise. The same cosmid libraries and selected cDNA library will be made available to all, and this will allow an efficient pooling of results. To this end a core database will be maintained by the coordinator and accessible by all participants through local copies of the same dataset. The complete network will be constructed with the IGD system, based on the ACEDB graphical database program. Large scale sequencing using state of the art technologies and strategies will be performed in areas where sequence ready maps are already available. This will be mainly performed in Xq28 which has been shown to contain an unusually high number of genes, many of which related to genetic disorders.

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Rue Laurent Fries 1

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Participants (6)