Lobo Neil F, Fraser Tresa S, Adams John A, Fraser Malcolm J
Department of Biological Sciences, Center for Tropical Diseases Research and Training, University of Notre Dame, PO Box 369, Notre Dame, IN 46556-0369, USA.
Genetica. 2006 Sep-Nov;128(1-3):347-57. doi: 10.1007/s10709-006-7165-2.
The piggyBac transposon is an extremely versatile helper-dependent vector for gene transfer and germ line transformation in a wide range of invertebrate species. Analyses of genome sequencing databases have identified piggyBac homologues among several sequenced animal genomes, including the human genome. In this report we demonstrate that this insect transposon is capable of transposition in primate cells and embryos of the zebrafish, Danio rerio. piggyBac mobility was demonstrated using an interplasmid transposition assay that has consistently predicted the germ line transformation capabilities of this mobile element in several other species. Both transfected COS-7 primate cells and injected zebrafish embryos supported the helper-dependent movement of tagged piggyBac element between plasmids in the characteristic cut-and-paste, TTAA target-site specific manner. These results validate piggyBac as a valuable tool for genetic analysis of vertebrates.
piggyBac转座子是一种极其通用的辅助依赖型载体，可用于多种无脊椎动物物种的基因转移和种系转化。对基因组测序数据库的分析已在包括人类基因组在内的多个已测序动物基因组中鉴定出piggyBac同源物。在本报告中，我们证明了这种昆虫转座子能够在斑马鱼（Danio rerio）的灵长类细胞和胚胎中发生转座。使用质粒间转座分析证明了piggyBac的移动性，该分析一直能够预测这种移动元件在其他几个物种中的种系转化能力。转染的COS-7灵长类细胞和注射的斑马鱼胚胎都支持标记的piggyBac元件以特征性的剪切粘贴方式在质粒之间进行辅助依赖型移动，且具有TTAA靶位点特异性。这些结果证实了piggyBac是用于脊椎动物遗传分析的有价值工具。
