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
Department of Molecular Genetics, The Ohio State University, Marion, OH 43302, USA.
Microbiology Program, The Ohio State University, Columbus, OH 43210, USA.
Methods Protoc. 2019 Aug 26;2(3):74. doi: 10.3390/mps2030074.
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.
准确的DNA损伤反应途径对于DNA双链断裂的修复至关重要。修复可通过同源重组发生，其中已鉴定出许多不同的子途径。一些重组途径是保守的，这意味着染色体序列得以保留，而其他途径是非保守的，会导致DNA序列发生一些改变。我们描述了一种体内遗传检测方法，用于研究……中非串联直接重复区域的非保守染色体内缺失。该检测方法可用于研究DNA复制过程中产生的自发断裂以及用同宗内切核酸酶（）产生的诱导双链断裂。该检测方法的初步遗传验证表明，自发断裂需要……但不需要……，而诱导断裂则需要这两个基因，这与先前的研究一致。该检测方法在DNA损伤修复领域对于研究染色体内缺失机制将是有用的。
