Insect Molecular Biology · 2000 · 130 citations · 39 references
Life SciencesEntomologyGeneticsGenetic EngineeringInsect GeneticsTransgenic CropsAgrobacterium-mediated TransformationPlant TransformationTransgene InheritanceTransgene StabilityTransgenesis StrategiesTransgenic Organism ProductionCotton Insect PestsGreen Fluorescent Protein ReportersMarker Gene ExpressionMarker Gene IntegrationSterile Insect TechniqueTransgenic InsectsTransposable Element ExpressionWestern Blot Analysis
The pink bollworm, Pectinophora gossypiella, is a world-wide pest of cultivated cotton. In certain growing regions populations are suppressed by a sterile release strategy. Efforts to improve the sterile insect technique as well as our understanding of lepidopteran biology could benefit greatly from a germ-line transformation system. We report transformation of pink bollworm with a piggyBac transposable element carrying the enhanced green flourescent protein (EGFP) marker gene. This vector-marker system resulted in recovery of transgenics at a rate of approximately 3.5%. Integration of the transforming construct that was typical of piggyBac was demonstrated by Southern analysis and sequence determination of transposon flanks. Expression of the EGFP marker was visualized by fluorescent microscopy and Western Blot analysis. Maintenance of transformed strains indicates that the transgene segregates in a Mendelian fashion and has been stable over fourteen generations to date.
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