ABSTRACT
We report the heritable germ-line transformation of the Queensland fruit fly, Bactrocera tryoni, using a piggyBac vector marked with either the fluorescent protein DsRed or EGFP. A transformation frequency of 5-10% was obtained. Inheritance of the transgenes has remained stable over more than 15 generations despite the presence of endogenous piggyBac sequences in the B. tryoni genome. The sequence of insertion sites shows the usual canonical pattern of piggyBac integraton into TTAA target sites. An investigation of endogenous piggyBac elements in the B. tryoni genome reveals the presence of sequences almost identical to those reported recently for the B. dorsalis complex of fruit flies and two noctuid moths, suggesting a common origin of piggyBac sequences in these species. The availability of transformation protocols for B. tryoni has the potential to deliver improvements in the performance of the Sterile Insect Technique for this pest species.
Subject(s)
Animals, Genetically Modified/genetics , DNA Transposable Elements/genetics , Genetic Vectors/genetics , Tephritidae/genetics , Transformation, Genetic , Animals , Female , Germ Cells , Male , Microinjections , Pest Control, BiologicalABSTRACT
Explant-derived cultured autologous epidermis was used as a graft in a 41-year-old female patient with burns, the first subject in a clinical trial of the technique. A small full-thickness biopsy specimen which was taken on Day 2 of the hospital admission was used to initiate epidermal cultures, four of which were grafted onto the patient's back and right leg 29 and 35 days later. Three of these epidermal cultures engrafted successfully, in spite of infection with Pseudomonas aeruginosa, which resulted in the loss of some of the conventional, split-thickness meshed autografts that were applied concurrently. The fourth graft, which may have been oriented incorrectly onto the graft bed, was largely unsuccessful, and only small islets of epithelial cells remained after 10 days. The successful grafts produced full-thickness, epidermal coverage with a good cosmetic result and little evidence of contraction during a six-months' follow-up.