In vivo biochemistry: Physical monitoring of recombination induced by site‐specific endonucleases

In vivo biochemistry: Physical monitoring of recombination induced by site‐specific endonucleases The recombinational repair of chromosomal double‐strand breaks (DSBs) is of critical importance to all organisms, who devote considerable genetic resources to ensuring such repair is accomplished. In Saccharomyces cerevisiae, DSB‐mediated recombination can be initiated synchronously by the conditional expression of two site‐specific endonucleases, HO or I‐Scel. DNA undergoing recombination can then be extracted at intervals and analyzed. Recombination initiated by meiotic‐specific DSBs can be followed in a similar fashion. This type of ‘in vivo biochemistry’ has been used to describe several discrete steps in two different homologous recombination pathways: gene conversion and single‐strand annealing. The roles of specific proteins during recombination can be established by examining DNA in strains deleted for the corresponding gene. These same approaches are now becoming available for the study of recombination in both higher plants and animals. Physical monitoring can also be used to analyze nonhomologous recombination events, whose mechanisms appear to be conserved from yeast to mammals. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png BioEssays Wiley

In vivo biochemistry: Physical monitoring of recombination induced by site‐specific endonucleases

BioEssays, Volume 17 (7) – Jul 1, 1995

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Publisher
Wiley
Copyright
Copyright © 1995 Cambridge University Press
ISSN
0265-9247
eISSN
1521-1878
D.O.I.
10.1002/bies.950170707
Publisher site
See Article on Publisher Site

Abstract

The recombinational repair of chromosomal double‐strand breaks (DSBs) is of critical importance to all organisms, who devote considerable genetic resources to ensuring such repair is accomplished. In Saccharomyces cerevisiae, DSB‐mediated recombination can be initiated synchronously by the conditional expression of two site‐specific endonucleases, HO or I‐Scel. DNA undergoing recombination can then be extracted at intervals and analyzed. Recombination initiated by meiotic‐specific DSBs can be followed in a similar fashion. This type of ‘in vivo biochemistry’ has been used to describe several discrete steps in two different homologous recombination pathways: gene conversion and single‐strand annealing. The roles of specific proteins during recombination can be established by examining DNA in strains deleted for the corresponding gene. These same approaches are now becoming available for the study of recombination in both higher plants and animals. Physical monitoring can also be used to analyze nonhomologous recombination events, whose mechanisms appear to be conserved from yeast to mammals.

Journal

BioEssaysWiley

Published: Jul 1, 1995

References

  • Mating‐type gene switching in Saccharomyces cerevisiae
    Haber, Haber
  • Genetic and physical analysis of double‐strand break repair and recombination in Saccharomyces cerevisiae
    Rudin, Rudin; Sugarman, Sugarman; Haber, Haber
  • Molecular analysis of V(D)J recombination
    Gellert, Gellert

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