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

Journal Article · Thu Dec 01 00:00:00 EST 1994 · Genomics

DOI:https://doi.org/10.1006/geno.1994.1656· OSTI ID:241034

Haldi, M ^[1]; Perrot, V; Saumier, M ^[2]

---

1. Whitehead Institute for Biomedical Research, Cambridge, MA (United States)
2. Centre d`Etude du Polymorphisms Humain, Paris (France); and others

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. 27 refs., 3 figs., 3 tabs.

• OSTI ID: 241034
• Journal Information: Genomics, Vol. 24, Issue 3; Other Information: PBD: Dec 1994
• Country of Publication: United States
• Language: English

Related Subjects

55 BIOLOGY AND MEDICINE
BASIC STUDIES
56 BIOLOGY AND MEDICINE
APPLIED STUDIES
99 MATHEMATICS
COMPUTERS
INFORMATION SCIENCE
MANAGEMENT
LAW
MISCELLANEOUS
HUMAN CHROMOSOMES
GENETIC RADIATION EFFECTS
MUTATION FREQUENCY
DNA-CLONING
CHIMERAS
DATA BASE MANAGEMENT
GENES
GENE RECOMBINATION
POLYMERASE CHAIN REACTION
YEASTS
FLUORESCENCE
DNA HYBRIDIZATION
RADIATION INDUCED MUTANTS