In the genome project, the first step of gene analysis has been the ongoing construction of contiguous genetic and physical maps and the large-scale accumulation of partial CDNA sequences to provide a working resource to the community. The next logical stage will be to integrate the gene and genome maps. Further research will then be needed to elucidate the functions of the encoded proteins, to unravel their potential role in genetic disease and to develop diagnostic tests and therapeutic approaches. The work of the first step is well under way in many places. This project aims to realise the next step, the integration of the gene and genome map by fluorescence in situ hybridisation (FISH). FISH technology is presently revolutionizing human cytogenetics and has strongly enhanced gene and genome mapping. We propose to apply existing and develop new FISH hybridization and microscopy technology to the genomic mapping of cdnas, for a rapid accumulation of positional information, at coarse and fine levels. Our approach consists of two, parallel strategies:
i) Direct, high-throughput multicolour FISH mapping of cdnas, without prior isolation of genomic clones. The research involves existing, but continually improving technology of (pro)metaphase FISH mapping using fluorescent counterstain banding. Emphasis will be put on increasing throughput by multicolour multiplexing, involving technology and instrumentation developed in parallel EC-projects on automated cytogenetics.
ii) High-resolution DNA-halo mapping and ordering of genes within genomic contigs from selected areas of interest. The Leiden groups have recently developed novel DNA-halo multicolour FISH technoloqy further increasing the resolution of FISH beyond that of interphase DNA. This technique will greatly propel in situ mapping at short-range (10-200 kb) and will be developed, as part of this project, for YAC-containing Yeast spheroplasts, for implementation by both groups to map genes in contigs of their interest, notably at 4q35, 16P13 and Xp22 for Leiden and 1p13, 11p13 and 12P13 for CNRS-Villejuif.
Funding SchemeCSC - Cost-sharing contracts