Long-read plasmid sequencing strategy for evaluating intrinsic instability of tandem gene arrays.

microPublication biology Pub Date : 2025-05-08 eCollection Date: 2025-01-01 DOI:10.17912/micropub.biology.001582
Hiroaki Takesue, Satoshi Okada, Takashi Ito
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Abstract

Tandem gene arrays are inherently unstable, particularly in recombination-prone organisms such as the budding yeast Saccharomyces cerevisiae . However, understanding the nature of this instability-how frequently and to what extent the target array contracts or expands within the genome-remains challenging. As a surrogate approach to this goal, we propose using nanopore long-read sequencing to directly determine the length distribution of a target gene array cloned onto a centromeric plasmid functioning as an artificial chromosome. This strategy will not only allow us to assess the intrinsic instability of the array but also help identify factors that may influence its stability.

评价串联基因阵列内在不稳定性的长读质粒测序策略。
串联基因阵列本身是不稳定的,特别是在易重组的生物体中,如出芽酵母酵母。然而,理解这种不稳定性的本质——目标阵列在基因组内收缩或扩展的频率和程度——仍然具有挑战性。作为实现这一目标的替代方法,我们建议使用纳米孔长读测序来直接确定克隆到作为人工染色体的着丝粒质粒上的靶基因阵列的长度分布。这种策略不仅使我们能够评估阵列的内在不稳定性,而且还有助于确定可能影响其稳定性的因素。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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