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Genome linking with yeast artificial chromosomes.

Coulson, A; Waterston, R; Kiff, J; Sulston, J; Kohara, Y

Nature. 1988;335(6186):184-6.

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Abstract

The haploid genome of Caenorhabditis elegans consists of some 80 x 10(6) base pairs of DNA contained in six chromosomes. The large number of interesting loci that have been recognized by mutation, and the accuracy of the genetic map, 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 together. 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 electrophoresis. 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) vector, 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.

Bibliographic metadata

Content type:
Published date:
Language:
eng
Journal title:
Abbreviated journal title:
ISSN:
Place of publication:
ENGLAND
Volume:
335
Issue:
6186
Pagination:
184-6
Digital Object Identifier:
10.1038/335184a0
Pubmed Identifier:
3045566
Access state:
Active

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Academic department(s):

Record metadata

Manchester eScholar ID:
uk-ac-man-scw:67571
Created by:
Green, Linda
Created:
22nd October, 2009, 09:11:29
Last modified by:
Green, Linda
Last modified:
12th November, 2012, 20:12:22

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