Abstract
The haploid genome of Caenorhabditis elegans consists of some 80 × 106 base pairs of DNA contained in six chromosomes1. The large number of interesting loci that have been recognized by mutation2, and the accuracy of the genetic map3, mean that a physical map of the genome is highly desirable, because it will facilitate the molecular cloning of chosen loci. The first steps towards such a map used a fingerprinting method to link cosmid clones together4. This approach reached its practical limit last year, when 90–95% of the genome had been cloned into 17,500 cosmids assembled into some 700 clusters (contigs), but the linking clones needed were either non-existent or extremely rare. Anticipating this, we had planned to link by physical means— probably by hybridization to NotI fragments separated by pulse field gel electrophoresis5–7. NotI recognizes an eight base sequence of GC pairs; thus the fragments should be large enough to bridge regions that clone poorly in cosmids, and, with no selective step involved, would necessarily be fully representative. However, with the availability of a yeast artificial chromosome (YAC) vector8, we decided to use this alternative source of large DNA fragments to obtain linkage. The technique involves the ligation of large (50–1,000 kilobase) genomic fragments into a vector that provides centromeric, telomeric and selective functions; the constructs are then introduced into Saccharomyces cerevisiae, and replicate in the same manner as the host chromosomes.
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Coulson, A., Waterston, R., Kiff, J. et al. Genome linking with yeast artificial chromosomes. Nature 335, 184–186 (1988). https://doi.org/10.1038/335184a0
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DOI: https://doi.org/10.1038/335184a0
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