Zheng Xiuzhong, Epstein Anastasiya, Klein Hannah L
Department of Biochemistry, New York University School of Medicine, New York, NY 10016, USA.
Methods Mol Biol. 2011;745:3-13. doi: 10.1007/978-1-61779-129-1_1.
Spontaneous mitotic recombination occurs in response to DNA damage incurred during DNA replication or from lesions that do not block replication but leave recombinogenic substrates such as single-stranded DNA gaps. Other types of damages result in general genome instability such as chromosome loss, chromosome fragmentation, and chromosome rearrangements. The genome is kept intact through recombination, repair, replication, checkpoints, and chromosome organization functions. Therefore when these pathways malfunction, genomic instabilities occur. Here we outline some general strategies to monitor a subset of the genomic instabilities: spontaneous mitotic recombination and chromosome loss, in both haploid and diploid cells. The assays, while not inclusive of all genome instability assays, give a broad assessment of general genome damage or inability to repair damage in various genetic backgrounds.
自发有丝分裂重组是对DNA复制过程中发生的DNA损伤或不阻碍复制但留下重组底物(如单链DNA缺口)的损伤作出的反应。其他类型的损伤会导致一般的基因组不稳定,如染色体丢失、染色体片段化和染色体重排。基因组通过重组、修复、复制、检查点和染色体组织功能保持完整。因此,当这些途径发生故障时,就会出现基因组不稳定。在这里,我们概述了一些监测基因组不稳定子集的一般策略:单倍体细胞和二倍体细胞中的自发有丝分裂重组和染色体丢失。这些检测方法虽然不包括所有的基因组不稳定检测,但能对各种遗传背景下的一般基因组损伤或无法修复损伤进行广泛评估。