The potato is the fifth most important crop in the world. Potato production in tropical and subtropical countries suffers from damage caused by the potato tuber moth, Phthorimaea operculella (PTM). Development of a germ line transformation system for the potato tuber moth will improve understanding of the basic biology of this insect and may reveal molecular targets for the development of new control strategies. To establish a robust system for PTM transformation, we tested three components that are critical to genetic transformation systems for insects; promoter activity, marker gene expression, and transposable element function. We compared the transcriptional activities of 5 different promoters, hsp70, hsp82, actin5C, polyubiquitin and IE1, within PTM embryos. The IE1 promoter flanked with the enhancer element, hr5, showed a very high level of transcriptional activity compared with all other tested promoters. The hr5-IE1 promoter was used to drive the expression of the enhanced green fluorescent protein (EGFP) as a marker gene. A high proportion of the embryos injected with the hr5-IE1/EGFP plasmid fluoresced under UV-illumination. The transpositional activities of the Hermes, mariner and piggyBac transposable elements were determined by inter-plasmid transposition assays. The observation of piggyBac transposition events enabled a transformation experiment to be designed using a piggyBac-hr5-IE1-EGFP vector.
Species 1: Lepidoptera Gelechiidae Phthorimaea operculella (potato tuber moth)
Keywords: Transposable elements, Enhanced green fluorescent protein (EGFP)
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