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Establishment of an RTA-Bddsx hybrid system for female-specific splicing that can affect the sex ratio of Bactrocera dorsalis (Hendel) after embryonic injection.

BACKGROUND
The oriental fruit fly, Bactrocera dorsalis (Hendel), a very destructive insect pest in many areas of Asia, including Taiwan, can cause significant damage by ovipositing in and larval feeding on many kinds of fruit. A female lethal system, combining the splicing property of doublesex (dsx) with the toxicity of ricin A chain (RTA), has been developed. In this system, a modified RTA is separated by Bddsx intron 3; the expressed RNA can only be spliced in females, with toxic effects, whereas the immature RTA in males is harmless.


RESULTS
Two RTA-Bddsx constructs, clone BE 24-7 and clone CF 26-21, containing Bddsx intron 3 and its flanking exonic sequences, with four nucleotides at the 5'-end and five nucleotides at the 3'-end, correctly spliced in a sex-specific manner. Wild-type and modified RTAs expressed in an Escherichia coli system retained their ability to suppress protein synthesis: 90.4% for Ricin-WT, 71.3% for Ricin-LERQ and 58.0% for Ricin-FEGQ. Embryonic injection of Acp-CF26-21, the RTA-Bddsx gene driven by the actin 5C promoter, resulted in a significant increase in male percentage in the eclosed adults.


CONCLUSION
Our results indicate that the RTA-Bddsx hybrid system offers a novel and promising approach for oriental fruit fly control.