Abstract
The tuberous sclerosis disease gene TSC1 has been mapped to 9q34. However, its precise localisation has proved problematic because of conflicting recombination data. Therefore, we have attempted to clone the entire target area into cosmid contigs prior to gene isolation studies. We have used Alu-PCR from irradiation hybrids to produce complex probes from the target region which have identified 1,400 cosmids from a chromosome-specific library. These, along with cosmids obtained by other methods, have been assembled into contigs by a fingerprinting technique. We estimate that we have obtained most of the region in cosmid contigs. These cosmids are a resource for the isolation of expressed genes within the TSC1 interval. In addition, the cosmid contig assembly has demonstrated a number of previously unknown physical connections between genes and markers in 9q34.
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Acknowledgements
The chromosome-specific gene library LL09NC01 ‘P’ used in this work was constructed at the Biomedical Sciences Division, Lawrence Livermore National Laboratory, Livermore, Calif., USA, under the auspices of the National Laboratory Gene Library Project sponsored by the US Department of Energy. We are grateful to Bart Janssen, Marjon van Slegtenhorst and colleagues for sharing their data in advance of publication. We would like to thank Carol Jones for providing hybrid line 64063a12 and A.M. Frischauf for providing the facilities at ICRF for replicating the LL09NC01 ‘P’ library, as well as spotting it onto filters. We thank M.W. Burley for technical assistance and Lynn West for help with the in situ hybridisation. We are grateful to Duncan Franklin for supplying us with some unpublished cosmid data. The irradiation and fusion procedure was carried out at ICRF under the supervision of Louise Sefton and Peter Goodfellow. J.N. and J.F. were supported by grants from the Tuberous Sclerosis Association of Great Britain. The work was also supported by grants from the MRC UK Human Genome Mapping Project.
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Nahmias, J., Hornigold, N., Fitzgibbon, J. et al. Cosmid Contigs Spanning 9q34 Including the Candidate Region for TSC1. Eur J Hum Genet 3, 65–77 (1995). https://doi.org/10.1159/000472279
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DOI: https://doi.org/10.1159/000472279
