|Ferguson, Holly -|
|Thibault, Steve -|
|Mohammed, Ahmed -|
|Fraser, Malcon -|
Submitted to: Insect Molecular Biology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: March 26, 2010
Publication Date: January 21, 2011
Citation: Ferguson, H.J., Neven, L.G., Thibault, S., Mohammed, A., Fraser, M. 2011. Genetic Transformation of the Codling Moth, Cydia pomonella L., with piggyBac EGFP. Insect Molecular Biology. 20:201-214. Interpretive Summary: Codling moth is a pest of apples, pears, and stone fruits grown in the United States. Scientists at the USDA-ARS laboratory in Wapato, Washington, Washington State University, Amgen, Inc. of California, and the University of Notre Dame collaborated to demonstrate that codling moth could be stably transformed with foreign DNA usiing a transposable element called piggyBac. This is the first description of successful genetic transformation of the codling moth. By stably transforming this insect, scientists proved that conditionally sterile lines could be developed for use in field suppression efforts similar to the sterile insect technique that is in use. Conditionally sterile transformed insects would be released into the field to mate with wild-type insects and bring about population suppression.
Technical Abstract: Genetic transformation of the codling moth, Cydia pomonella, was accomplished through embryo microinjection with a plasmid-based piggyBac vector containing the enhanced green fluorescent protein (EGFP) gene. Sequencing of the flanking regions around the inserted construct results in identification of insect genomic sequences, not plasmid sequences, thus providing evidence that the piggyBac EGFP cassette had integrated into the codling moth genome. EGFP-positive moths were confirmed as long as 28 generations post injection through PCR and Southern analyses, indicating heritability of the transgene.