Washington State University, 24106 North Bunn Road, Prosser, WA 99350, USA.
Transgenic Res. 2011 Feb;20(1):201-14. doi: 10.1007/s11248-010-9391-8. Epub 2010 Apr 13.
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 resulted 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 in the 28th and earlier generations post injection through PCR and Southern blot analyses, indicating heritability of the transgene.
通过用含有增强型绿色荧光蛋白(EGFP)基因的基于质粒的 piggyBac 载体对苹果蠹蛾胚胎进行显微注射,实现了苹果蠹蛾的遗传转化。对插入构建体周围侧翼区域的测序导致鉴定出昆虫基因组序列,而不是质粒序列,从而提供了 piggyBac EGFP 盒已整合到苹果蠹蛾基因组中的证据。通过 PCR 和 Southern blot 分析,在注射后的第 28 代及更早世代中确认了 EGFP 阳性的蛾,表明转基因的可遗传性。