Screening and evaluation of promoters for effective expression of fluorescent protein and high performance of the modified hyPBase in Zeugodacus cucurbitae

Abstract

The melon fly (Zeugodacus cucurbitae(Coquillett)) is a highly destructive quarantine pest that inflicts significant damage on the agricultural industry. However, the absence of efficient genetic research tools for this pest hinders its research progress and therefore its control. Among these, the selection of an appropriate promoter plays a central role. In this study, the activities of numerous promoter systems were examined by transfecting Drosophila S2 cells and microinjecting Z. cucurbitae embryos. The results reveal that native promoters drive higher expression within their respective species than in others and that the ZcPUb promoter effectively drives the expression of fluorescent proteins in Z. cucurbitae. A hyperactive piggyBac transposase (hyPBase) was also genetically modified to the control of the ZcPUb promoter and it significantly improved the transgenesis efficiency for constructs of different sizes. The temporal and spatial expression patterns of fluorescence regulated by the ZcPUb promoter in transgenic flies suggest that the ZcPUb promoter-controlled fluorescence can be used as a valuable marker for transgenic screening of Z. cucurbitae. These findings not only highlight the ZcPUb promoter as a crucial resource for future investigations in gene functions and developmental processes in Z. cucurbitae, but also emphasize the importance of identifying native promoters for research and practical application of organisms.