LARGE HUMAN YACS CONSTRUCTED IN A RAD52 STRAIN SHOW A REDUCED RATE OF CHIMERISM

被引：37

作者：

HALDI, M

PERROT, V

SAUMIER, M

DESAI, T

COHEN, D

CHERIF, D

WARD, D

LANDER, ES

机构：

[1] WHITEHEAD INST BIOMED RES,CAMBRIDGE CTR 9,CAMBRIDGE,MA 02142

[2] FDN JEAN DAUSSET,CTR ETUD POLYMORPHISME HUMAIN,F-75010 PARIS,FRANCE

[3] YALE UNIV,SCH MED,DEPT GENET,NEW HAVEN,CT 06520

[4] MIT,DEPT BIOL,CAMBRIDGE,MA 02139

来源：

GENOMICS | 1994年 / 24卷 / 03期

关键词：

D O I：

10.1006/geno.1994.1656

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

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. (C) 1994 Academic Press, Inc.

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页码：478 / 484

页数：7

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