Germ-line transformation of pink bollworm (Lepidoptera: gelechiidae) mediated by the piggyBac transposable element.
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.
Peloquin
J J
JJ
Department of Entomology, UC Riverside, Riverside, CA, peloquin@faculty.ucr.edu
Thibault
S T
ST
Staten
R
R
Miller
T A
TA
D000818:Animals; D001483:Base Sequence; D004251:DNA Transposable Elements; D005260:Female; D005822:Genetic Vectors; D005854:Germ Cells; D049452:Green Fluorescent Proteins; D015151:Immunoblotting / Q000379:methods; D008164:Luminescent Proteins / Q000235:genetics; D008297:Male; D008845:Microinjections / Q000379:methods; D008969:Molecular Sequence Data; D009036:Moths / Q000235:genetics*; D013194:Staining and Labeling / Q000379:methods; D014170:Transformation, Genetic
D004251:DNA Transposable Elements; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins