Gabriel Abram, Dapprich Johannes, Kunkel Mark, Gresham David, Pratt Stephen C, Dunham Maitreya J
Department of Molecular Biology and Biochemistry, Rutgers University, Piscataway, New Jersey, United States of America.
PLoS Genet. 2006 Dec 15;2(12):e212. doi: 10.1371/journal.pgen.0020212. Epub 2006 Nov 1.
Transposable genetic elements are ubiquitous, yet their presence or absence at any given position within a genome can vary between individual cells, tissues, or strains. Transposable elements have profound impacts on host genomes by altering gene expression, assisting in genomic rearrangements, causing insertional mutations, and serving as sources of phenotypic variation. Characterizing a genome's full complement of transposons requires whole genome sequencing, precluding simple studies of the impact of transposition on interindividual variation. Here, we describe a global mapping approach for identifying transposon locations in any genome, using a combination of transposon-specific DNA extraction and microarray-based comparative hybridization analysis. We use this approach to map the repertoire of endogenous transposons in different laboratory strains of Saccharomyces cerevisiae and demonstrate that transposons are a source of extensive genomic variation. We also apply this method to mapping bacterial transposon insertion sites in a yeast genomic library. This unique whole genome view of transposon location will facilitate our exploration of transposon dynamics, as well as defining bases for individual differences and adaptive potential.
可移动遗传元件无处不在,然而在基因组内任何给定位置上它们的存在与否在不同的单个细胞、组织或菌株之间可能会有所不同。可移动元件通过改变基因表达、协助基因组重排、导致插入突变以及作为表型变异的来源,对宿主基因组产生深远影响。鉴定基因组中全部转座子需要进行全基因组测序,这使得对转座作用对个体间变异影响的简单研究变得不可能。在此,我们描述了一种用于确定任何基因组中转座子位置的全局定位方法,该方法结合了转座子特异性DNA提取和基于微阵列的比较杂交分析。我们使用这种方法来绘制酿酒酵母不同实验室菌株中内源性转座子的图谱,并证明转座子是广泛基因组变异的一个来源。我们还将此方法应用于绘制酵母基因组文库中细菌转座子插入位点。这种独特的转座子位置全基因组视图将有助于我们探索转座子动态,以及确定个体差异和适应潜力的基础。