Department of Genetics, University of Leicester, University Road, Leicester LE17RH, UK.
Mol Biotechnol. 2011 Mar;47(3):243-52. doi: 10.1007/s12033-010-9338-6.
Intracisternal A-type particle (IAP) elements are high copy number long terminal repeat (LTR) rodent retrotransposons. Some IAP elements can transpose, and are responsible for ~12% of spontaneous mouse mutations. Inbred mouse strains show variation in genomic IAP distribution, contributing to inter-strain genetic variability. Additionally IAP elements can influence the transcriptional regulation of neighbouring genes through their strong LTR promoter, effecting phenotypic variation. This genetic and phenotypic variability can translate into experimental variability between mouse strains. For example, it has been demonstrated that strain-specific genetic/epigenetic factors can interact to yield variable responses to drugs. Therefore, in experimental contexts it is essential to unequivocally identify mouse strains. Recently it was estimated that any two inbred strains share only ~40% of their IAP insertions. Of the remaining 60%, some insertions will be strain specific, fixed during inbreeding. These fixed insertions can be exploited as genetic markers to identify inbred strains, if they can be identified simply and efficiently. Here, we report the development of a PCR-based system allowing direct acquisition of strain-specific IAP insertions. In a pilot study, we identified 21 IAP loci, genotyped IAP insertions at 9 loci, and discovered two strain-specific insertions that could reliably identify these strains.
内含 A 型粒子 (IAP) 元件是高拷贝数长末端重复 (LTR) 啮齿动物逆转录转座子。一些 IAP 元件可以转座,这是约 12%的自发突变的原因。近交系小鼠品系的基因组 IAP 分布存在差异,导致了种间遗传变异性。此外,IAP 元件可以通过其强大的 LTR 启动子影响邻近基因的转录调控,从而影响表型变异。这种遗传和表型的可变性可能转化为不同小鼠品系之间的实验可变性。例如,已经证明,特定于品系的遗传/表观遗传因素可以相互作用,从而对药物产生不同的反应。因此,在实验背景下,明确识别小鼠品系至关重要。最近估计,任何两个近交系仅共享约 40%的 IAP 插入。在剩下的 60%中,一些插入将是种特异性的,在近交过程中固定。如果这些固定的插入可以简单有效地被识别,它们可以作为遗传标记来识别近交系。在这里,我们报告了一种基于 PCR 的系统的开发,该系统允许直接获取种特异性的 IAP 插入。在一项试点研究中,我们确定了 21 个 IAP 基因座,对 9 个基因座的 IAP 插入进行了基因分型,并发现了两个可以可靠识别这些品系的种特异性插入。