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Contenu archivé le 2024-04-16

A deletion, restriction site and overlapping clone map of the human Y chromosome

Objectif

The project is aimed at creating a physical map of the human Y chromosome by bringing together several physical mapping approaches and resources available to the collaborating laboratories. These will include deletion mapping using patients with structural abnormalities of the Y; pulsed-field gel electrophoresis analysis using Y-specific probes; development of overlapping clone sets of the Y based on clones isolated from YAC and cosmid libraries.
An overlapping clone map, based on yeast artificial chromosome (YAC) clones and cosmid clones is being constructed for the human Y chromosome.
The research has been successful in exploiting the resources that the collaborating groups were able to bring to bare on the development of a more detailed map of the human Y chromosome. The deletion maps of both the short and long arms of the Y chromosome. The detail will be increased further as more patients and markers are contributed to the analysis. The YAC map has also been expanded which gives a high level of overlapping coverage of the Y euchromatin. This coverage is further increased by combination with YAC clones. Detailed verification of the YAC contigs defined by sequence tagged site (STS) content analysis is underway by developing restriction maps of individual clones and by Alu fingerprint analysis. Further detailed restriction analysis has been carried out of YACs covering selected regions of the map. This is being extended to the rest of the euchromatin. Cosmid contigs which are estimated to cover 69% of the Y chromosome are being developed through Hinf I fingerprint analysis.
The main tasks are:

The development and integration of the participants' existing deletion maps of Yp and Yq which are based on analysis of patient material with Y-specific DNA probes. To add new patients and probes to this map and to integrate those studied by others into a unitary deletion map of the Y.

The construction of long range restriction maps of each deletion interval by pulsed-field gel electrophoresis and, eventually, linking these together.

The use of probes in each deletion interval to isolate large clones from YAC and cosmid libraries currently available and to assemble these into overlapping clone sets for each interval.

The development of an extended overlapping clone map by linking deletion intervals together. A further large insert YAC library from the OXEN cell line (49,XYYYY) will be constructed. This library will be generally available.

The analysis of overlapping clone sets defining deletion intervals for protein coding sequences beginning with those where genetic functions have been mapped through karyotype to phenotype correlations in patients.

This approach, based on deletion intervals where a large number of Y probes have already been assigned, will provide the basis for the establishment of an extensive physical map of the Y chromosome as an important step towards complete closure.

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Coordinateur

University of Cambridge
Contribution de l’UE
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Adresse
Tennis Court Road
CB2 1QP Cambridge
Royaume-Uni

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