Nucleic Acids Research, 1987, Vol. 15, No. 4 1363-1375
© 1987
Articles |
Progress towards construction of a total restriction fragment map of a human chromosome
1Department of Biochemistry, St. Mary's Hospital Medical School, University of London London W2 1PG, UK 2Hagedorm Research Laboratory Niels Steensensvej 6, DK-2820 Gentofte, Denmark 3Division of Molecular Genetics, National Institute for Medical Research The Ridgeway, Mill Hill, London NW7 4Imperial Cancer Research Fund, Lincoln's Inn Fields London WC2 5Metabolic Unit, St. Mary's Hospital, London W2, UK
Received January 6, 1987. Accepted January 21, 1987.
We present an approach to the construction of an overlapping restriction fragment map of a single human chromosome. A genomic cosmid library genome was constructed from a mouse-human hybrid cell line containing chromosome 17 as its only human genetic component. Cosmids containing human inserts were isolated by hybridisation to a human Alu sequence. DNAs from ninety-six randomly chosen cosmids were digested with either EcoRI or HindIII, end-labelled with 35S-dATP and analysed using agarose gel electrophoresis. Comparison of the restriction fragment patterns revealed two pairs of overlapping clones, that were confirmed by cross-hybridisation of the overlapping fragments. The two pairs of cosmids both mapped to human chromosome 17, as shown by hyb ridisation to a panel of somatic cell hybrids. These data demonstrate that the generation of an overlapping cosmid map along a human chromosome is feasible, representing an intermediate step towards the complete sequencing of a human chromosome.