Project Number: 6615-22000-025-04
Start Date: Sep 01, 2009
End Date: Aug 31, 2014
The genetic components for these systems will be first identified and isolated from either A. ludens or A. suspensa. These will include regulatory DNA sequences from embryonic genes such as serendipity, slam, and mdg88; sex-specific intron splicing cassettes isolated from the A. ludens sex-determination gene transformer; and the lethal effector gene, hid. To test the Mediterranean fruit fly embryonic lethality system in Anastrepha species existing driver (srya-tTA) and effector (TRE-hidala5) transgene constructs shown to be functional in the Mediterranean fruit fly will be integrated into a wild A. ludens host strain. Endogenous early and late embryonic genes will be isolated from the Anastrepha species to test their promoters in driver constructs, and efforts will be made to isolate the hid lethal-effector from Anastrepha. Transgenic A. suspensa and A. ludens will be created using the strongest early and late promoter-tra intron-tTA constructs (based on tTA transcription) inserted into DsRed-marked piggyBac vector plasmids having the TRE-tra intron-hidala5 driver and phiC31 attP integrase recombination sites. Transformation experiments will be initiated in A. ludens using transgene vectors yielding the most efficient female-lethality in A. suspensa. If no homozygous line yields 100% female-lethality, the transgenes will be re-mobilized by transposase injection to create new insertion sites that will be tested for more optimal parameters. Optimal homozygous lines will be expanded for large scale sexing, fitness, and mating competitiveness tests in Tapachula and Petapa.