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An Assay to Study Intra-Chromosomal Deletions in Yeast

1
Department of Molecular Genetics, The Ohio State University, Marion, OH 43302, USA
2
Microbiology Program, The Ohio State University, Columbus, OH 43210, USA
3
Molecular Genetics Program, The Ohio State University, Columbus, OH 43210, USA
4
Department of Molecular Genetics, The Ohio State University, Columbus, OH 43210, USA
*
Author to whom correspondence should be addressed.
Methods Protoc. 2019, 2(3), 74; https://doi.org/10.3390/mps2030074
Received: 6 August 2019 / Revised: 20 August 2019 / Accepted: 21 August 2019 / Published: 26 August 2019
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 Schizosaccharomyces pombe. This assay can be used to study both spontaneous breaks arising during DNA replication and induced double-strand breaks created with the S. cerevisiae homothallic endonuclease (HO). The preliminary genetic validation of this assay shows that spontaneous breaks require rad52+ but not rad51+, 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. View Full-Text
Keywords: DNA double-strand breaks; Genetic Recombination; Yeast DNA double-strand breaks; Genetic Recombination; Yeast
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MDPI and ACS Style

Lucas, B.E.; McPherson, M.T.; Hawk, T.M.; Wilson, L.N.; Kroh, J.M.; Hickman, K.G.; Fitzgerald, S.R.; Disbennett, W.M.; Rollins, P.D.; Hylton, H.M.; Baseer, M.A.; Montgomery, P.N.; Wu, J.-Q.; Petreaca, R.C. An Assay to Study Intra-Chromosomal Deletions in Yeast. Methods Protoc. 2019, 2, 74. https://doi.org/10.3390/mps2030074

AMA Style

Lucas BE, McPherson MT, Hawk TM, Wilson LN, Kroh JM, Hickman KG, Fitzgerald SR, Disbennett WM, Rollins PD, Hylton HM, Baseer MA, Montgomery PN, Wu J-Q, Petreaca RC. An Assay to Study Intra-Chromosomal Deletions in Yeast. Methods and Protocols. 2019; 2(3):74. https://doi.org/10.3390/mps2030074

Chicago/Turabian Style

Lucas, Bailey E.; McPherson, Matthew T.; Hawk, Tila M.; Wilson, Lexia N.; Kroh, Jacob M.; Hickman, Kyle G.; Fitzgerald, Sean R.; Disbennett, W. M.; Rollins, P. D.; Hylton, Hannah M.; Baseer, Mohammed A.; Montgomery, Paige N.; Wu, Jian-Qiu; Petreaca, Ruben C. 2019. "An Assay to Study Intra-Chromosomal Deletions in Yeast" Methods Protoc. 2, no. 3: 74. https://doi.org/10.3390/mps2030074

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