Mutation burst by conflict of two mouse subspecies genomes

两个小鼠亚种基因组冲突导致突变爆发

• 批准号: 23657009
• 负责人: SHIROISHI Toshihiko
• 金额: $ 2.58万
• 依托单位: National Institute of Genetics
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Challenging Exploratory Research
• 财政年份: 2011
• 资助国家: 日本
• 起止时间: 2011 至 无数据
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-23657009/
• 关键词: ゲノム構築; 再編; 維持; 遺伝学; ゲノム; 進化; 変異誘導

We carried out two experiments to elucidate mechanism by which high-frequency spontaneous mutations(Recombination-induced-mutation : Rim) is induced in mice with recombinant MHC haplotypes between two mouse subspecies. First, we tried to detect genome alterations similar to Rim by PCR-based monitoring of Rim-type mutations using pooled sperm DNAs of intra-MHC recombinant mice. As a result, we could not observe expected PCR-products by this method. One possibility to explain this result is that there are different types of mechanism for induction of Rim. Alternatively, present intra-MHC recombinant mice have lost the activity to give rise to Rim. Second, we tried to develop an efficient assay system to detect DNA-double strand breaks and subsequent recombination for single male mouse. To do this, now we constructed the DNA construct that consists of recombination-target DNA sequence for Prdm9 protein, which is known to determine recombination site by introducing Histon3 chemical modification as hotspot mark, and of GFP reporter connected to the Prdm9 target sequence. Now we are introducing this construct into mouse genome by means of Tol2 transposon system.

我们进行了两个实验，以阐明机制，通过这些机制，在两种小鼠亚种之间具有重组MHC单倍型的小鼠中，诱导了高频自发突变（重组引起的突变：RIM）。首先，我们试图通过使用MHC内部重组小鼠的合并精子DNA对RIM型突变进行基于PCR的RIM型突变的监测来检测类似于RIM的基因组改变。结果，我们无法通过这种方法观察到预期的PCR产品。解释这一结果的一种可能性是，有不同类型的RIM诱导机制。或者，当前的MHC内重组小鼠已经失去了活性，从而引起边缘。其次，我们试图开发一个有效的测定系统，以检测单一雄性小鼠的DNA双链断裂和随后的重组。为此，现在我们构建了由PRDM9蛋白重组 - 目标DNA序列组成的DNA构建体，该蛋白已知可以通过将Histon3化学修饰作为热点标记以及连接到PRDM9目标序列连接的GFP Reporter来确定重组位点。现在，我们通过TOL2转座子系统将该构建体引入小鼠基因组。

项目成果

Prdm9は種分化遺伝子か?-野生マウスの多型から考える-

Prdm9 是物种形成基因吗？ - 考虑到野生小鼠的多态性 -

• 发表时间: 2011
• 作者: 河野宏光,城石俊彦;河野宏光,田村勝,長田直樹,鈴木仁,太田邦史,城石俊彦
• 通讯作者: 河野宏光,田村勝,長田直樹,鈴木仁,太田邦史,城石俊彦

生物の科学遺伝: Review,相同染色体間組換えのホットスポット-研究の歴史と最近の急展開

生物体的科学遗传学：同源染色体间重组的回顾、热点——研究历史和近期快速发展

• 发表时间: 2012
• 期刊: 日本語総説(エヌ・ティー・エス社)
• 作者: 河野宏光,城石俊彦