shotgun cloning of transposon insertions in the genome of caenorhabditis elegans猎枪克隆转座子插入在秀丽隐杆线虫的基因组.pdf

Comparative and Functional Genomics Comp Funct Genom 2004; 5: 225–229. Published online in Wiley InterScience (). DOI: 10.1002/cfg.392 Short Communication Shotgun cloning of transposon insertions in the genome of Caenorhabditis elegans Alexander M. van der Linden and Ronald H. A. Plasterk* Hubrecht Laboratory, Uppsalalaan 8, 3584 CT, Utrecht The Netherlands *Correspondence to: Abstract Ronald H. A. Plasterk, Hubrecht Laboratory, Uppsalalaan 8, 3584 We present a strategy to identify and map large numbers of transposon insertions in CT, Utrecht, The Netherlands. the genome of Caenorhabditis elegans. Our approach makes use of the mutator strain E-mail: plasterk@niob.knaw.nl mut-7, which has germline-transposition activity of the Tc1/mariner family of trans- posons, 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 Received: 15 April 2003 of C. elegans mutants. Copyright  2004 John Wiley Sons, Ltd. Revised: 21 January 2004 Keywords: Caenorhabditis elegans ; insertional mutagenesis screen; mut-7 ; Tc1; Accepted: 28 January 2004 transposon insertion display protocol Introduction 1996). In the genomic sequence of the commonly used Bristol N2 C.