Lorenzen M D, Berghammer A J, Brown S J, Denell R E, Klingler M, Beeman R W
Division of Biology, Kansas State University, Manhattan, KS, USA.
Insect Mol Biol. 2003 Oct;12(5):433-40. doi: 10.1046/j.1365-2583.2003.00427.x.
The lepidopteran transposable element piggyBac can mediate germline insertions in at least four insect orders. It therefore shows promise as a broad-spectrum transformation vector, but applications such as enhancer trapping and transposon-tag mutagenesis are still lacking. We created, cloned, sequenced and genetically mapped a set of piggyBac insertions in the red flour beetle, Tribolium castaneum. Transpositions were precise, and specifically targeted the canonical TTAA recognition sequence. We detected several novel reporter-expression domains, indicating that piggyBac could be used to identify enhancer regions. We also demonstrated that a primary insertion of a non-autonomous element can be efficiently remobilized to non-homologous chromosomes by injection of an immobile helper element into embryos harbouring the primary insertion. These developments suggest potential for more sophisticated methods of piggyBac-mediated genome manipulation.
鳞翅目转座元件piggyBac可介导至少四个昆虫目的种系插入。因此，它有望成为一种广谱转化载体，但诸如增强子捕获和转座子标签诱变等应用仍很缺乏。我们在赤拟谷盗（Tribolium castaneum）中创建、克隆、测序并对一组piggyBac插入进行了遗传定位。转座是精确的，并且特异性靶向典型的TTAA识别序列。我们检测到了几个新的报告基因表达结构域，表明piggyBac可用于鉴定增强子区域。我们还证明，通过将一个不能移动的辅助元件注射到含有初次插入的胚胎中，一个非自主元件的初次插入可以有效地转移到非同源染色体上。这些进展表明了piggyBac介导的基因组操作更复杂方法的潜力。
