Large human YACs constructed in a rad52 strain show a reduced rate of chimerism.

Genomics
Authors
Keywords
Abstract

Current YAC libraries are plagued by a high frequency of chimeras--that is, clones containing fragments from multiple genomic regions. Chimeras are thought to arise largely through recombination in the yeast host cell. If so, the use of recombination-deficient yeast strains, such as rad52 mutants, might be expected to alleviate the problem. Here, we report the construction of megabase-sized human YACs in the rad52 strain MHY5201 and the determination of their rate of chimerism by fluorescence in situ hybridization analysis. Examination of 48 YACs showed a rate of chimerism of at most 8%, whereas YACs constructed in the wildtype host AB1380 showed a rate of about 50%. These results show that it is possible to significantly decrease the rate of YAC chimerism through the use of appropriate yeast host strains.

Year of Publication
1994
Journal
Genomics
Volume
24
Issue
3
Pages
478-84
Date Published
1994 Dec
ISSN
0888-7543
DOI
10.1006/geno.1994.1656
PubMed ID
7713499
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