An Assay to Study Intra-Chromosomal Deletions in Yeast

An accurate DNA damage response pathway is critical for the repair of DNA double-strand breaks. Repair may occur by homologous recombination, of which many different sub-pathways have been identified. Some recombination pathways are conservative, meaning that the chromosome sequences are preserved,...

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Bibliographic Details
Published in:Methods and protocols 2019-08, Vol.2 (3), p.74
Main Authors: Lucas, Bailey E, McPherson, Matthew T, Hawk, Tila M, Wilson, Lexia N, Kroh, Jacob M, Hickman, Kyle G, Fitzgerald, Sean R, Disbennett, W Miguel, Rollins, P Daniel, Hylton, Hannah M, Baseer, Mohammed A, Montgomery, Paige N, Wu, Jian-Qiu, Petreaca, Ruben C
Format: Article
Language:English
Subjects:
DNA double-strand breaks
Genetic Recombination
Yeast
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Summary:An accurate DNA damage response pathway is critical for the repair of DNA double-strand breaks. Repair may occur by homologous recombination, of which many different sub-pathways have been identified. Some recombination pathways are conservative, meaning that the chromosome sequences are preserved, and others are non-conservative, leading to some alteration of the DNA sequence. We describe an in vivo genetic assay to study non-conservative intra-chromosomal deletions at regions of non-tandem direct repeats in . This assay can be used to study both spontaneous breaks arising during DNA replication and induced double-strand breaks created with the homothallic endonuclease ( ). The preliminary genetic validation of this assay shows that spontaneous breaks require but not , while induced breaks require both genes, in agreement with previous studies. This assay will be useful in the field of DNA damage repair for studying mechanisms of intra-chromosomal deletions.
ISSN:2409-9279
2409-9279
DOI:10.3390/mps2030074