ABSTRACT
The absence of gene vectors for germline transformation of non-drosophilid insects remains a major barrier to both applied and basic advances in the molecular genetic analysis of insects. Routine and efficient gene transformation of insects is limited to Drosophila melanogaster, relying primarily on P element-based gene vectors. Functionality of the transposase gene in the host embryo is determined by hobo excision from the indicator plasmid, which deletes the lacZ reporter gene. A lack of excision maintains the lacZ reporter gene resulting in ß-galactosidase activity in transformed bacteria, which results in blue colonies on X-gal media. Both precise and imprecise hobo excisions from the indicator plasmid result in the loss of lacZ, yielding white bacterial colonies. The data indicate that cross-mobilizable systems, perhaps related to hobo, exist in a broad range of drosophilids. Hobo excision assays were performed in several tephritid species using the pKHFLIacZ indicator plasmid and the pHSH2 helper.
