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10.3390/mps2030074
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Article

An Assay to Study Intra-Chromosomal Deletions in Yeast

by
Bailey E. Lucas
1,
Matthew T. McPherson
1,
Tila M. Hawk
1, 1,
Jacob M. Kroh
1,
Kyle G. Hickman
1,
Sean R. Fitzgerald
1, 2,
P. Daniel Rollins
3,
Hannah M. Hylton
1,
Mohammed A. Baseer
1,
Paige N. Montgomery
1,
Jian-Qiu Wu
4 and
Ruben C. Petreaca
1,*
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
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Abstract

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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