Department of Biochemistry and Cellular and Molecular Biology, University of Tennessee, Knoxville, TN, USA.
UT-ORNL Graduate School of Genome Science and Technology, University of Tennessee, Knoxville, TN, USA.
Methods Mol Biol. 2021;2369:251-261. doi: 10.1007/978-1-0716-1681-9_14.
Two distinct groups of parasitic nematodes use programmed DNA elimination to silence germline-expressed genes in the somatic cells (ascarids) or for sex determination (Strongyloides spp.). In the ascarids, DNA is lost only in pre-somatic cells during early embryogenesis, leading to a reduced somatic genome compared to the intact germ cell genome. Comparative genome analysis has provided information on the retained vs. eliminated sequences, DNA breaks, a full chromosome view on DNA elimination, and the evolutionary conservation of DNA elimination among ascarids. These studies have revealed novel insights into the functions and mechanisms of DNA elimination and provided a reference for in-depth molecular analysis of DNA elimination. Here, I describe the genomics methods we used to study programmed DNA elimination, focusing on the parasitic nematode Ascaris.
两组不同的寄生性线虫利用程序性 DNA 消除来使生殖细胞表达的基因在体细胞中沉默(蛔虫)或用于性别决定(旋毛虫属)。在蛔虫中,只有在早期胚胎发生过程中的前体细胞中才会丢失 DNA,导致与完整的生殖细胞基因组相比,体细胞基因组减少。比较基因组分析提供了有关保留和消除序列、DNA 断裂、DNA 消除的完整染色体视图以及蛔虫中 DNA 消除的进化保守性的信息。这些研究揭示了 DNA 消除的功能和机制的新见解,并为 DNA 消除的深入分子分析提供了参考。在这里,我将描述我们用于研究程序性 DNA 消除的基因组学方法,重点介绍寄生性线虫蛔虫。