Requirements for ectopic homologous recombination in mammalian somatic cells

Mark D. Baker; Leah R. Read; Barbara G. BeattyPhilip Ng

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Requirements for ectopic homologous recombination in mammalian somatic cells

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Ectopic recombination occurs between DNA sequences that are not in equivalent positions on homologous chromosomes and has beneficial as well as potentially deleterious consequences for the eukaryotic genome. In the present study, we have examined ectopic recombination in mammalian somatic (murine hybridoma) cells in which a deletion in the mu gene constant (Cmu) region of the endogenous chromosomal immunoglobulin mu gene is corrected by using as a donor an ectopic wild- type Cmu region. Ectopic recombination restores normal immunoglobulin M production in hybridomas. We show that (i) chromosomal mu gene deletions of 600 bp and 4 kb are corrected less efficiently than a deletion of only 2 bp, (ii) the minimum amount of homology required to mediate ectopic recombination is between 1.9 and 4.3 kb, (iii) the frequency of ectopic recombination does not depend on donor copy number, and (iv) the frequency of ectopic recombination in hybridoma lines in which the donor and recipient Cmu regions are physically connected to each other on the same chromosome can be as much as 4 orders of magnitude higher than it is for the same sequences located on homologous or nonhomologous chromosomes. The results are discussed in terms of a model for ectopic recombination in mammalian somatic cells in which the scanning mechanism that is used to locate a homologous partner operates preferentially in cis.

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《Molecular and Cellular Biology》 |1996年第12期|7122-7132|共11页
作者
Mark D. Baker; Leah R. Read; Barbara G. BeattyPhilip Ng;
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Departments of Pathobiology University of Guelph, Guelph, Ontario, Canada N1G 2W1;

Department of Pediatric Laboratory Medicine, Hospital for Sick Children, Toronto, Ontario, Canada M5G 1X8;

Molecular Biology and Genetics, University of Guelph, Guelph, Ontario, Canada N1G 2W1;

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正文语种英语
中图分类分子生物学;
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Requirements for ectopic homologous recombination in mammalian somatic cells

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