Assessment of Ubiquitous Promoters Driving Fluorescent Marker and Transposase Expression to Develop a...

Bactrocera dorsalis (oriental fruit fly) is a destructive invasive pest threatening global agriculture. Although integrated pest management is applied, environmentally friendly genetic control methods are urgently needed. The development of such methods particularly relies on efficient genetic elements. In this study, we compared the transient expression of mScarlet-I driven by various Actin and PUb promoters in B. dorsalis embryos. The truncation of two strong promoters, BdActA3a and BdPUb, revealed that the 5.0-kb BdActA3a and 3.6-kb BdPUb promoters drove significantly higher expression than their truncated variants. Notably, the BdPUb promoter was highly effective in driving fluorescent protein expression in B. dorsalis. Using the 3.6-kb BdPUb promoter, we constructed a transposase plasmid BdPUb-3.6 kb>hyPBase. By co-injecting the BdPUb 3.6kb>mScarlet-I donor construct, we successfully generated a fluorescent transgenic strain with a transgenic efficiency of approximately 26%. The strain exhibited stage-specific fluorescence and maternal effect and the homozygotes showed fecundity comparable to wild-type controls. The high performance of the piggyBac transposase and the fluorescence screening system provides a substantial technical foundation for basic research and future development of genetically modified strains to control B. dorsalis.