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Yeast artificial chromosomes: an alternative approach to the molecular analysis of mouse developmental mutations

Published online by Cambridge University Press:  14 April 2009

Zoia Larin  and
Hans Lehrach
Zoia Larin
Affiliation:
Imperial Cancer Research Laboratories, 44 Lincoln's Inn Fields, London WC2A 3PX
Hans Lehrach*
Affiliation:
Imperial Cancer Research Laboratories, 44 Lincoln's Inn Fields, London WC2A 3PX
*
* Corresponding author.
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Mammalian genetics now allows a molecular study of genomic regions previously analysed by genetic and embryological techniques. To simplify such an analysis, we have established a number of libraries of mouse DNA in Yeast Artificial Chromosome (YAC) vectors, constructed either by partial digestion with EcoRI, or by complete digestion with enzymes which cut rarely in the mammalian genome. In this paper we report the construction of complete digest libraries prepared from mouse genomic DNA using the rare cutter enzymes NoiI and BssHII, and the detection of gene loci from the H-2 complex, the t–complex, and other loci from the mouse genome. Due to their large insert size, YAC clones simplify the cloning of extended regions of the mouse genome surrounding known developmental mutations and should, after introduction into the germ line, offer a high probability of correct expression of the genes contained within the cloned region. We hope that this will allow the use of YAC clones to scan regions of interest such as the t–complex for specific genes by testing DNA introduced into transgenic mice for the ability to complement mutations localised to this region.

Type
Research Article
Information
Genetics Research , Volume 56 , Issue 2-3 , October 1990 , pp. 203 - 208
Copyright
Copyright © Cambridge University Press 1990

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