Fraser M J, Ciszczon T, Elick T, Bauser C
Department of Biological Sciences, University of Notre Dame, Indiana, 46556, USA.
Insect Mol Biol. 1996 May;5(2):141-51. doi: 10.1111/j.1365-2583.1996.tb00048.x.
Transposon mutagenesis of baculoviruses provides an ideal experimental system for analysis of the movement of a unique family of mobile element identified from lepidopteran genomes. Members of this family of short-inverted-repeat elements are characterized by their extreme specificity for TTAA target sites. This report describes the analysis of excision events for two representatives of this family, tagalong (formerly TFP3) and piggyBac (formerly IFP2). These elements were tagged with a polyhedrin/lacZ reporter gene and inserted back into the virus genome either by homologous recombination or by transposition. Revertants were selected based on a white plaque phenotype. Both elements excise in a precise fashion from their positions in the baculovirus genome in either TN-368 cells or IPLB-SF21 AE cells. The precise excision of these elements in infected IPLB-SF21 AE cells occurs in the absence of either tagalong or piggyBac element encoded functions. The common characteristics of both insertion and excision for these elements provides further validation for their inclusion in a single family of unique transposons.
杆状病毒的转座子诱变提供了一个理想的实验系统,用于分析从鳞翅目基因组中鉴定出的一类独特移动元件的移动情况。这个短反向重复元件家族的成员其特点是对TTAA靶位点具有极高的特异性。本报告描述了对该家族的两个代表,即“搭便车者”(原TFP3)和“piggyBac”(原IFP2)的切除事件分析。这些元件用多角体蛋白/乳糖酶Z报告基因进行标记,然后通过同源重组或转座重新插入病毒基因组。基于白色噬菌斑表型筛选回复突变体。在TN - 368细胞或IPLB - SF21 AE细胞中,这两种元件都能从杆状病毒基因组中的位置精确切除。在受感染的IPLB - SF21 AE细胞中,这些元件的精确切除在没有“搭便车者”或“piggyBac”元件编码功能的情况下也会发生。这些元件插入和切除的共同特征为将它们归入一个独特的转座子单一家族提供了进一步的验证。