Peloquin J J, Thibault S T, Staten R, Miller T A
Department of Entomology, UC Riverside, Riverside, CA,
Insect Mol Biol. 2000 Jun;9(3):323-33. doi: 10.1046/j.1365-2583.2000.00194.x.
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.
棉红铃虫(Pectinophora gossypiella)是一种危害全球棉花种植的害虫。在某些种植区域,通过不育释放策略来抑制其种群数量。建立种系转化系统将极大地有助于改进不育昆虫技术以及我们对鳞翅目生物学的理解。我们报道了利用携带增强型绿色荧光蛋白(EGFP)标记基因的piggyBac转座元件对棉红铃虫进行转化。这种载体 - 标记系统产生转基因个体的比率约为3.5%。通过Southern分析和转座子侧翼序列测定,证实了piggyBac典型的转化构建体的整合。通过荧光显微镜和蛋白质免疫印迹分析观察到EGFP标记的表达。转化品系的维持表明转基因以孟德尔方式分离,并且至今在十四代中一直保持稳定。