Progress towards construction of a total restriction fragment map of a human chromosome.
Nucleic Acids Research , Volume 15 p. 1363- 1375
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-hybridization of the overlapping fragments. The two pairs of cosmids both mapped to human chromosome 17, as shown by hybridization 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.
|*Chromosomes, Human, Pair 17, *DNA Restriction Enzymes, 0 (Cosmids), Animals, Cell Line, Chromosome Mapping, Cosmids, EC 3.1.21 (DNA Restriction Enzymes), Human, Hybrid Cells/cytology, Mice, Support, Non-U.S. Gov't|
|Nucleic Acids Research|
|Organisation||Erasmus MC: University Medical Center Rotterdam|
Vissing, H, Grosveld, F.G, Solomon, E, Moore, G, Lench, N, Shennan, N, & Williamson, R. (1987). Progress towards construction of a total restriction fragment map of a human chromosome. Nucleic Acids Research, 15, 1363–1375. Retrieved from http://hdl.handle.net/1765/2403