Abstract
We present a rapid in vitro method to scan the repair of DNA double-strand breaks (DSBs). A DSB was introduced at the EcoRI site within the lacZ gene of the plasmid pUC18 and the plasmid was exposed to cellular extracts from a wild-type repair-competent (RAD) and a mutant (rad52Δ) strain of the yeast Saccharomyces cerevisiae. The fidelity of rejoining was determined by the expression of the lacZ gene after bacterial transformation with the treated plasmid. A cellular extract from the yeast S. cerevisiae was found to be capable of rejoining DNA DSBs. Breaks at the EcoRI site were rejoined by extracts from both wild-type and mutant strains to form circular plasmids with almost equal efficiency. However, the fidelity of rejoining was lower for the rad52Δ extract than for normal wild-type.
Similar content being viewed by others
Author information
Authors and Affiliations
Additional information
Received: 2 September /2 November 1997
Rights and permissions
About this article
Cite this article
Jha, B., Ahne, F., Kistler, M. et al. A rapid method to monitor repair and mis-repair of DNA double-strand breaks by using cell extracts of the yeast Saccharomyces cerevisiae. Curr Genet 33, 1–3 (1998). https://doi.org/10.1007/s002940050300
Issue Date:
DOI: https://doi.org/10.1007/s002940050300