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A rapid method to monitor repair and mis-repair of DNA double-strand breaks by using cell extracts of the yeast Saccharomyces cerevisiae |
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| Authors: | Bhavanath Jha F Ahne Manfred Kistler Christian Klaus Friederike Eckardt-Schupp |
| Institution: | Department of Biotechnology, L. N. Mithila University, Darbhanga-846 008, India, IN Institut für Strahlenbiologie, GSF – Forschungszentrum für Umwelt und Gesundheit GmbH, Neuherberg, Postfach 1129, D-85758 Oberschlei?heim, Germany Fax: +49-3187 3381 e-mail: Ahne@GSF.DE, DE |
| 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. Received: 2 September /2 November 1997 |
| Keywords: | Yeast cellular extract DNA double-strand break Fidelity of repair |
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