Abstract
The piggyBac transposable element, originally isolated from a virus in an insect cell line, is a valuable molecular tool for transgenesis and mutagenesis of invertebrates. For heterologous transgenesis in a variety of mammals, transfer of the piggyBac transposable element from an ectopic plasmid only requires expression of piggyBac transposase. To determine if piggyBac could function in dicotyledonous plants, a two-element system was developed in tobacco (Nicotiana tabacum) to test for transposable element excision and insertion. The first transgenic line constitutively expressed piggyBac transposase, while the second transgenic line contained at least two non-autonomous piggyBac transposable elements. Progeny from crosses of the two transgenic lines was analyzed for piggyBac excision and transposition. Several progeny displayed excision events, and all the sequenced excision sites exhibited evidence of the precise excision mechanism characteristic of piggyBac transposase. Two unique transposition insertion events were identified that each included diagnostic duplication of the target site. These data indicate that piggyBac transposase is active in a dicotyledonous plant, although at a low frequency.
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Acknowledgments
We thank Malcom Fraser for supplying the piggyBac transposable element and transposase. We are grateful to Keith Johnson and Stephen Hughes for comments on a draft manuscript. David Lee, Mark Doehring, and Kay Shopinski provided excellent technical assistance. This study was supported by CRIS Project 3620-42000-040-00D base funds from the USDA. The mention of firm names or trade products in this manuscript does not imply that they are endorsed or recommended by the USDA over other firms or similar products not mentioned. USDA is an equal opportunity provider and employer.
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Communicated by S. Hohmann.
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Johnson, E.T., Dowd, P.F. A non-autonomous insect piggyBac transposable element is mobile in tobacco. Mol Genet Genomics 289, 895–902 (2014). https://doi.org/10.1007/s00438-014-0860-2
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DOI: https://doi.org/10.1007/s00438-014-0860-2