Project Number: 6615-22000-016-18
Project Type: Trust

Start Date: Sep 15, 2002
End Date: Sep 14, 2005

Objective:
To determine the phlylogenetic distribution of piggyBac in the Bactrocera, Spodoptera, and other insect species in several insect orders. To test the presence of functional piggyBac elements or cross-mobilizing systems in species harboring piggyBac by embryonic mobility assays, and to isolate these elements as genomic DNA. To create and test piggyBac vector constructs that can be immobilized after genomic integration by rearranging or deleting sequences required for mobility. To test the use chemical selection markers with fluorescent protein visible markers for mass selection and verification of transformant insects.

Approach:
The phlylogenetic distribution of piggyBac in the Bactrocera, Spodoptera, and other insect species will be determined by PCR gene amplification methods and screening of genomic libraries. The presence of functional elements in these species will be tested by embryonic transposition and excision assays with and without exogenous transposase, and functionality of isolated piggyBac and related elements will be tested by plasmid-based mobility assays and germ-line transformation experiments. New piggyBac vectors will be created that can be immobilized after genomic integration by use of the FRT/FLP recombination system so that vector sequences critical to mobility will be deleted or rearranged after chromosomal integration. New vectors will also be marked with chemical and drug resistance genes (neomycin-phosphotransferase, dieldrin-resistance, and organophosphorus dehydrogenase) and green (GFP) and red (DsRed) fluorescent protein markers so that transgenic insects can be mass-screened by chemcial selection and then verified by a visible marker. BSL-1; June 18, 2001.

Project Team

Handler, Alfred - Al

Project Annual Reports

FY 2003

Related National Programs

Plant, Microbial & Insect Genetic Res., Genomics, & Genetic Improv. I (301)   Crop Protection & Quarantine (304)