秀丽隐杆线虫基因组中转座子插入的鸟枪法克隆
Shotgun cloning of transposon insertions in the genome of Caenorhabditis elegans.
作者信息
van der Linden Alexander M, Plasterk Ronald H A
机构信息
Hubrecht Laboratory, Uppsalalaan 8, Utrecht 3584 CT, The Netherlands.
出版信息
Comp Funct Genomics. 2004;5(3):225-9. doi: 10.1002/cfg.392.
Abstract
We present a strategy to identify and map large numbers of transposon insertions in the genome of Caenorhabditis elegans. Our approach makes use of the mutator strain mut-7, which has germline-transposition activity of the Tc1/mariner family of transposons, a display protocol to detect new transposon insertions, and the availability of the genomic sequence of C. elegans. From a pilot insertional mutagenesis screen, we have obtained 351 new Tc1 transposons inserted in or near 219 predicted C. elegans genes. The strategy presented provides an approach to isolate insertions of natural transposable elements in many C. elegans genes and to create a large-scale collection of C. elegans mutants.
摘要
我们提出了一种在秀丽隐杆线虫基因组中识别和定位大量转座子插入的策略。我们的方法利用了突变菌株mut-7，它具有Tc1/水手家族转座子的种系转座活性、一种用于检测新转座子插入的展示方案以及秀丽隐杆线虫的基因组序列。通过初步的插入诱变筛选，我们在219个预测的秀丽隐杆线虫基因中或其附近获得了351个新的Tc1转座子。所提出的策略提供了一种方法，可用于分离许多秀丽隐杆线虫基因中的天然转座元件插入，并创建一个大规模的秀丽隐杆线虫突变体集合。
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