Finding the Mouse Disorganization Gene

描述（由申请人提供）：组织解体（Ds）是一种对小鼠发育具有深远影响的单基因突变。与大多数突变不同，具有诊断特征的特定性状在几代人之间遗传，具有Ds突变的小鼠的可遗传性状是以不可预测的方式产生出生缺陷的倾向-没有两只小鼠以相同的方式受到影响。受影响的小鼠表现出发育不全、畸形或结构和器官的重复。所有的解剖特征都容易发生畸形。值得注意的是，这些小鼠不容易患癌症，斑点，行为，长寿或生育问题。我们以前表明，Ds是一个真正的显性，获得功能（或显性负）突变的哺乳动物中的少数几个例子之一。确定Ds基因的身份是重要的，因为可以深入了解发育过程中模式形成的调控。此外，我们可以深入了解突变的分子本质，这些突变显示出可变表达和低突变率，这是人类许多出生缺陷的特征。 我们将Ds定位到小鼠Chr 14的0.2 cM（约600 kb）片段上，克隆到两个BAC重叠群（129/Sv和C57 BL/6 J）中，获得了约260 kb的野生型基因组序列，鉴定了约20个候选基因，并通过序列分析消除了2个基因，通过重组消除了7个基因，以及其他4个候选基因的大部分外显子作为Ds突变位点。 基于这些结果，我们现在提出了两个具体目标：具体目标1：通过完成Ds/Ds突变纯合子的Ds基因座的基因组序列并将其与野生型序列进行比较来识别候选Ds突变。具体目标2：检测打了就跑的工程突变小鼠中候选DNA序列的差异。

DESCRIPTION (provided by applicant): Disorganization (Ds) is a single gene mutation that has profound effects on mouse development. Unlike most mutations where particular traits with diagnostic features are inherited among generations, the heritable trait in mice with the Ds mutation is the propensity to make birth defects in an unpredictable manner - no two mice are affected in identical ways. Affected mice show agenesis, malformation, or duplication of structures and organs. All anatomical features are prone to malformation. Remarkably, these mice are not prone to cancer, spotting, behavioral, longevity or fertility problems. We previously showed that Ds is one of the few examples in mammals of a true dominant, gain-of-function (or dominant negative) mutation. Determining the identity of the Ds gene is important because deep insights into the regulation of pattern formation during development can be gained. In addition, we may gain insights into the molecular nature of mutations that show variable expression and low penetrance that are characteristic of many birth defects in humans. We mapped Ds to a 0.2 cM (about 600 kb) segment of mouse Chr 14, cloned in two BAC contigs (129/Sv and C57BL/6J), obtained about 260 kb of wild-type genomic sequence, identified about 20 candidate genes, and eliminated two genes by sequence analysis, seven genes by recombination, and most of the exons of four other candidate genes as the site of the Ds mutation. Based on these results, we now propose two Specific Aims: Specific Aim 1: To identify candidate Ds mutations by completing the genomic sequence of the Ds locus from Ds/Ds mutant homozygotes and comparing this to the wild-type sequence. Specific Aim 2: To test candidate DNA sequence differences in hit-and-run engineered mutant mice.