In Vivo Construction of Transgenes in Drosophila

摘要: Transgenic flies are generated by transposon-mediated transformation. A drawback of this approach is the size limit transposable elements. Here, we propose a novel method that allows extension transgenes in vivo. This based on an incomplete transgene has been constructed vitro and integrated into Drosophila genome conventional transgenesis. The contains two short stretches DNA homologous to 5′- 3′-ends larger segment interest. Between homology I-SceI recognition site located. Once activated, endonuclease introduces double-strand break, which triggers ectopic recombination between endogenous locus. Through gap repair, obtains complete region interest Our results show effective for copying up 28 kb genomic transgene, thereby eliminating technical difficulties associated with construction large extending limits current transgenesis protocols. In general, may be useful technique genetic engineering eukaryotic model organisms.