Large human YACs constructed in a rad52 strain show a reduced rate of chimerism.
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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
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Journal | Genomics
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Volume | 24
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Issue | 3
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Pages | 478-84
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Date Published | 1994 Dec
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ISSN | 0888-7543
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DOI | 10.1006/geno.1994.1656
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PubMed ID | 7713499
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